DNA Analysis Unit II Quality Assurance Manual

Transcription

1 DNA Analysis Unit II Quality Assurance Manual DNA Analysis Unit II Quality Assurance Manual 2.doc Issue Date: 03/26/08 Revision: 2 Page 1 of 20 DNA Analysis Unit II Quality Assurance Manual 1 Mission, Goals, and Objectives 1.1 Mission Statement To conduct forensic mitochondrial DNA (mtdna) examinations of biological evidence in criminal, terrorism, and missing persons cases to support the FBI and other law enforcement agencies; and to provide leadership to the forensic mtdna community through training, population and missing person databasing, regional mtdna laboratory analysis services, and research or validation of new technologies. 1.2 Goals and Objectives The goals of DNAU II include: Conduct mtdna examinations of biological evidence to support federal, state and local law enforcement agencies, to include evidence processing, report writing, and expert testimony Hire and train additional personnel to perform mtdna analysis in order to reach the DNAU II target staffing level, to reduce the examiner caseload to fewer than 30 cases and to improve turnaround time for mtdna cases to meet FBI Laboratory-established guidelines Validate new technologies regarding mtdna analysis and implement these technologies into forensic casework. Specifically, new technologies should focus on improved DNA recovery, faster analysis, and more informative analysis of the mtdna genome Encourage DNAU II examiners and biologists to continue to publish scientific articles regarding mtdna analysis in order to facilitate exchange of mtdna information and technology transfer Provide liaison with other governmental, university and private laboratories regarding mtdna analysis through the mtdna subcommittee and the Combined DNA Index System (CODIS) subcommittee of the Scientific Working Group on DNA Analysis Methods (SWGDAM), the National Missing Person DNA Database (NMPDD) Program, and the mtdna Database Program. This is a controlled document.

2 DNA Analysis Unit II Quality Assurance Manual DNA Analysis Unit II Quality Assurance Manual 2.doc Issue Date: 03/26/08 Revision: 2 Page 2 of Manage four regional mtdna laboratories for the purpose of conducting no-cost forensic mtdna analysis to state and local law enforcement agencies in the United States and its territories Educate state and local forensic laboratory personnel about mtdna analysis for possible implementation of mtdna technology to other state and local forensic laboratories through the two week AForensic mtdna Analysis@ course at the FBI Academy and the FBI Laboratory. This course is accredited for three graduate hours at the University of Virginia Educate prosecutors and law enforcement agencies about mtdna analysis and its usefulness in criminal investigations to assist in homicide investigations, missing persons cases and in presenting mtdna evidence in court Implement mtdna analysis into the National DNA Index System (NDIS) in the form of an unidentified human remains index and a relatives of missing persons index, using Combined DNA Index System (CODIS) software The objectives of the Quality Assurance Program for DNAU II are to continuously improve the overall quality of DNA testing services by: Routinely monitoring Unit procedures through quality control standards, proficiency testing, and audits Confirming that the analytical data and interpretation of DNA testing results are scientifically accurate, and presented in a manner of high quality Establishing and maintaining a system for documenting procedures and policies within DNAU II. 2 Organization The organization of DNAU II is described below. The available positions within the Unit are the Unit Chief, Program Manager(s), Biologist-Forensic Examiner (Examiner), Biologist, and Program Analyst. All technical personnel meet the requirements of the FBI Quality Assurance Standards for Forensic DNA Testing Laboratories (FBI Standards). 2.1 Unit Chief DNAU II is headed by a Unit Chief. The Unit Chief is the Technical Leader of the Unit and is responsible for overall coordination of DNAU II programs, budget, monthly statistics, case assignments, liaison, implementation of new technologies, hiring, training, administrative review This is a controlled document.

3 DNA Analysis Unit II Quality Assurance Manual DNA Analysis Unit II Quality Assurance Manual 2.doc Issue Date: 03/26/08 Revision: 2 Page 3 of 20 of casework, proficiency testing, ensuring adherence to FBI Laboratory policies and practices for quality assurance, various personnel issues, and supervising all unit members. 2.2 Program Manager A Program Manager is responsible for each program administered by DNAU II. A Program Manager may also serve as an Examiner in a program. The programs in the Unit include Research and Validation, Training, Population Database, Quality Assurance (QA), Indian Country, National Missing Person DNA Database, mtdna Operations, TEDAC, JFAIDD, Regional Mitochondrial DNA Laboratories, and Mass Spectrometry. 2.3 Examiners and Biologists Casework can be conducted by Examiners and Biologists working together, or solely by Examiners. If a team approach is used, then, generally, a team consists of one Examiner and one or more Biologists. The Examiner supervises the Biologist(s) on his or her team. Biologists carry out laboratory procedures from DNA extraction through DNA sequencing, while Examiners conduct sequence analysis and interpretation, generate reports and provide expert testimony. One or more Biologists may function as Training Biologists and Quality Assurance (QA) Biologists, but also can carry out laboratory procedures on casework on a limited basis. 3 Authority and Accountability for the Quality Assurance Program The Quality Assurance Program of DNAU II is organized as follows: 3.1 Unit Chief The DNAU II Unit Chief is ultimately responsible for quality assurance activities within DNAU II. The DNAU II Unit Chief is responsible for ensuring the completeness of laboratory notes, reports, and casework and database documentation, ensuring that all Examiners and Biologists receive the necessary training and are qualified for their assigned work responsibilities, approving the use of analytical methods within DNAU II and establish criteria for procedural validation, and ensuring that corrective action is taken and documented when appropriate. The DNAU II Unit Chief serves as the Technical Leader of the unit. 3.2 QA Program Manager The QA Program Manager is appointed by the Unit Chief and is responsible for ensuring that all aspects of the DNAU II Quality Assurance Program are within FBI Laboratory guidelines and in compliance with the FBI Standards, for reviewing and maintaining quality assurance and quality control documentation to include proficiency testing records, and for developing, updating, and This is a controlled document.

4 DNA Analysis Unit II Quality Assurance Manual DNA Analysis Unit II Quality Assurance Manual 2.doc Issue Date: 03/26/08 Revision: 2 Page 4 of 20 implementing procedures within the Quality Assurance Program regarding FBI Laboratory policies and practices and DNAU II policies and procedures. The QA Program Manager is responsible for providing training to the DNAU II relevant to the policies, practices, and procedures within the Quality Assurance Program. 3.3 QA Biologists The QA Biologists are responsible for identifying specific problems in the Unit requiring quality assurance monitoring, for planning and determining reagent, instrumentation, and equipment needs of DNAU II, evaluating same, and providing recommendations to the Unit Chief with supporting documentation necessary for decision making, and for solving technical and quality control problems and improving the methods and processes carried out in DNAU II. 4 Job Descriptions, Training, and Continuing Education The Unit Chief, Examiners, and Biologists meet the requirements of the FBI Laboratory Quality Assurance Manual, Section and the FBI Standards. 4.1 Job Descriptions Job descriptions for the following positions in DNAU II are maintained in the FBI Laboratory unit responsible for maintaining human resource information. 4.2 Training $ Unit Chief GS-15 $ Program Manager(s) GS-14 $ Biologist-Forensic Examiner GS-7/9/11/12/13/14 $ Biologist GS-5/7/9/11/12/13 $ Program Analyst GS-12/13 DNAU II complies with Sections , , and of the FBI Laboratory Quality Assurance Manual. Training of Examiners and Biologists who perform mtdna analysis is in accordance with the DNAU II Examiner Training Program and Biologist Training Program, respectively. The training records and certificates of qualification are maintained in the Unit. The Unit Chief maintains documentation on the completion of training for each Examiner and Biologist. 4.3 Continuing Education DNAU II complies with Section of the FBI Laboratory Quality Assurance Manual. In This is a controlled document.

5 DNA Analysis Unit II Quality Assurance Manual DNA Analysis Unit II Quality Assurance Manual 2.doc Issue Date: 03/26/08 Revision: 2 Page 5 of 20 addition, the Technical Leader and each Examiner must stay abreast of developments within the field of DNA typing in accordance with the FBI Standards. Continuing education requirements of the FBI Standards are fulfilled by reading current scientific literature, as well as by attending seminars, courses, professional meetings or documented training sessions/classes in relevant subject areas at least once a year. Training needs are assessed by the Unit Chief during the mid-year Performance Reviews and the end-of-year Performance Appraisals. The effectiveness of the continuing education is evaluated by discussion between the employee and supervisor, and is documented by each of them placing his/her initials and date of discussion at the bottom of the Continuing Education Program Certification, form FD-932. Annual records of continuing education are kept by each person in the form of a Statement of Qualifications, and a copy maintained by the Unit Chief. 5 Security of Facilities DNAU II complies with Section 3.8 of FBI Laboratory Quality Assurance Manual, the FBI Laboratory Operations Manual (LOM) Practices for Laboratory Security, and the Security Program Reference Guide for Laboratory Division Personnel. The following outlines specific security measures taken by DNAU II. 5.1 Laboratory and Office Access Each Unit employee is assigned a key for accessing the Unit's exterior office doors. Unit laboratory space is accessed by the employee badge and laboratory key. Access to DNAU II office and laboratory space is limited to FBI Laboratory employees. Access to Rooms 3191 and 3399, the main evidence storage rooms for DNAU II, is limited to DNAU II employees, designated Evidence Control Unit employees, and a limited number of facilities maintenance employees and is accessed by a combination of an employee's badge and individual personal identification numbers. These rooms are alarmed when not occupied by DNAU II employees. 5.2 Evidence Storage Areas Each Examiner is assigned a refrigerator for storage of evidence in Room 3191 or Keys are maintained by Examiners and/or Biologists in a designated area A duplicate set of keys to evidence storage areas is maintained in the Unit Tel-Key cabinets in Room DNAU II employees have access to the secondary evidence storage areas (refrigerators and freezers) in Rooms 3181 and These refrigerators and freezers are locked during non-work hours. The secondary evidence stored in these areas is from cases under active examination. This is a controlled document.

6 DNA Analysis Unit II Quality Assurance Manual DNA Analysis Unit II Quality Assurance Manual 2.doc Issue Date: 03/26/08 Revision: 2 Page 6 of Key control lists are maintained in the Unit. 5.3 Closing of Unit At the end of each work day, each Examiner or Biologist will ensure that the evidence storage for the Examiner is locked. The last person to leave the Unit's laboratory space will ensure that the Unit's laboratory doors are locked. 6 Environmental Health and Safety All laboratory operations will be performed in a safe manner and in accordance with the standards established by applicable regulatory agencies. DNAU II complies with Section 3.4 of the FBI Laboratory Quality Assurance Manual and the FBI Laboratory Safety Manual. The FBI Laboratory Safety Manual is available to employees on the FBI Laboratory intranet. The Unit has a designated Safety Officer. A representative from the Health and Safety Group (HSG) serves as the FBI Laboratory Safety Officer. 7 DNAU II Documentation 7.1 Responsibility The QA Program Manager ensures that appropriate DNAU II quality assurance documentation is maintained. 7.2 Documentation and Records Maintained by DNAU II $ Unit Quality Assurance Manual $ Mitochondrial DNA Analysis Protocol $ Equipment Inventory $ Chemical Inventory $ Solution Preparation Records $ Quality Control Records for Critical Reagents $ Audit Records $ Relevant Safety Records $ Proficiency Testing Records $ Court Testimony Monitoring Records $ Unit Training Documentation $ Continuing Education Records $ Key Control Lists This is a controlled document.

7 7.3 Document Control $ Equipment Calibration Schedules/Records $ Equipment Maintenance Schedules/Records DNA Analysis Unit II Quality Assurance Manual DNA Analysis Unit II Quality Assurance Manual 2.doc Issue Date: 03/26/08 Revision: 2 Page 7 of 20 The DNAU II Mitochondrial DNA Analysis Protocol, the DNAU II Training Manual, the DNAU II Quality Assurance Manual, and one copy of each instrument manual for the ABI Prism 3130 Genetic Analyzers, the Gene Amp Systems 9700, and the Agilent 2100 Bioanalyzers are subject to the LOM Practices for Document Control and are prepared in accordance with these practices. The DNAU II Quality Assurance Manual and the DNAU II Mitochondrial DNA Analysis Protocol are available to Unit employees on the FBI Laboratory intranet. Controlled copies of the DNAU II Mitochondrial DNA Analysis Protocol are distributed to the Unit Chief, as well as to Examiners, and Biologists who perform the procedure(s). Controlled copies of the Training Manual are distributed to Examiner and Biologist trainees, and one is maintained by the QA Program Manager. The official version of the DNAU II Quality Assurance Manual and the DNAU II Mitochondrial DNA Analysis Protocol is located on the FBI Laboratory Intranet. The controlled documents listed in the previous paragraph are reviewed annually by the Unit Chief. These reviews are documented. An archived copy of each superseded controlled document is maintained in the Unit. 8 Evidence Control DNAU II conforms with Sections 3.12 and 5.8 of the FBI Laboratory Quality Assurance Manual and the LOM Practices for Laboratory Security, Practices for Recording and Acknowledging Evidence, Practices for Inventorying and Identifying Evidence, Practices for Processing a Request for Examination, Practices for the Examination of Evidence, Practices for Shipping Evidence, Practices for Transferring Evidence, Practices for Handling Drug and Valuable Evidence and Practices for Securing Evidence Storage Rooms regarding evidence control. DNAU II complies with Section 5.7 of the FBI Laboratory Quality Assurance Manual regarding evidence sampling. Section 3 of the Mitochondrial DNA Analysis Protocol describes evidence sampling in detail. The following outlines specific evidence control measures taken by DNAU II. It is noted that while every attempt is made not to consume the entirety of any particular item of evidence to allow for possible reexamination of that evidence at a later date by another laboratory, the primary goal is to use the amount of evidence necessary to provide mtdna typing results. 8.1 Evidence Receipt for mtdna Casework Evidence is received by the Examiner or Biologist assigned to the case from the designated Evidence Control Unit (ECU) employee. Typically, the ECU employee places the evidence in a This is a controlled document.

8 DNA Analysis Unit II Quality Assurance Manual DNA Analysis Unit II Quality Assurance Manual 2.doc Issue Date: 03/26/08 Revision: 2 Page 8 of 20 designated evidence storage area in the Unit. The Examiner or Biologist retrieving the evidence indicates its retrieval from Unit storage with a notation of such (for example, Unit Storage or DNAU II Storage ) in the Signature box of the Delivered By block on the first line of a FBI Laboratory Chain-of-Custody Log, form and/or 7-243a, and proper completion of the Accepted By block. Further evidence labeling and sealing/unsealing is performed in accordance with the relevant FBI Laboratory evidence control practices. 8.2 Evidence Storage for mtdna Casework Evidence such as bodily fluids, blood and saliva, hairs, bones, and teeth is generally stored refrigerated (typically 4 C ). With the exception of liquid blood, the above evidence may be stored frozen (typically -20 C) if deemed necessary. All DNA extracts and amplified DNA products are stored refrigerated (typically 4 C). Other evidence such as slides and pillboxes may be stored at room temperature. 8.3 Secondary Evidence Secondary evidence is defined as DNA extracts (and dilutions thereof), and powdered bone and tooth material with associated reagent blank swabs. Each DNA-containing tube is wrapped in Parafilm. Tubes are labeled with the item identifier and FBI Laboratory number and are packaged in sealed plastic bags. Bags with extracted DNA are labeled DNA Extracts. Typically, all secondary evidence is returned to ECU or to the contributor and is accompanied by the DNAU II Secondary Evidence Inventory (Appendix A). For cases where samples are entered into the NMPDD, a portion of each relevant sample and/or extract is retained and archived for possible reexamination. 8.4 Active Examination Evidence, whether primary or secondary, is considered to be under active examination, and, hence, subjected to provisions in the LOM for evidence under active examination, from the time the first sample in a submission (or a group of submissions in a case) is cut for DNA extraction until the time the Examiner determines that all laboratory analysis is complete for that submission or group of submissions. Typically, this time period does not exceed a few weeks. Once laboratory analysis is complete, evidence is stored properly sealed as defined by the applicable practices in the LOM. 8.5 Drug and Valuable Evidence Drug and valuable evidence received in DNAU II is handled in accordance with the LOM This is a controlled document.

9 DNA Analysis Unit II Quality Assurance Manual DNA Analysis Unit II Quality Assurance Manual 2.doc Issue Date: 03/26/08 Revision: 2 Page 9 of 20 Practices for Handling Drug and Valuable Evidence. 8.6 Evidence Return Evidence is transferred to the designated ECU employee who is responsible for ensuring the evidence is packaged and returned to the contributor. This may be accomplished by transferring the evidence to unit storage and notifying the designated ECU employee that the evidence is ready for pick up, or the evidence may be taken to ECU directly. 9 Validation New procedures used for mtdna analysis are developed and validated in accordance with Sections and of the FBI Laboratory Quality Assurance Manual, the LOM Practices for Developing Technical Procedures, the LOM Practices for Validating Technical Procedures, and the FBI Standards. Validation records conducted by DNAU II or by the Counterterrorism and Forensic Science Research Unit (CTFSRU) are kept in the Unit or at the Forensic Science Research and Training Center in Quantico, Virginia. Validated procedures are subjected to technical review and Unit Chief approval prior to implementation. Documentation of the review and approval is maintained in the Unit. The Unit retains copies of all relevant publications of validation studies conducted by the FBI Laboratory and by other laboratories. Modifications of procedures are documented as major or minor deviations. Major deviations are maintained in the unit and the relevant case files, and minor deviations are maintained in the relevant case files. 10 Standard Operating Procedures The Mitochondrial DNA Analysis Protocol describes the methods and procedures used in extracting, amplifying, and sequencing mtdna. This protocol is in conformance with the LOM Practices for Writing Standard Operating Procedures and Practices for Document Control. Any changes to the Mitochondrial DNA Analysis Protocol are done in compliance with the Section 5.4 of the FBI Laboratory Quality Assurance Manual and the LOM Practices for Writing Standard Operating Procedures and Practices for Document Control prior to implementation. Document approval records are maintained in the Unit and in QATU. 11 Case Assignment, Documentation and Review All Examiners and Biologists who perform casework document all aspects of the analysis processes in accordance with Sections 3.9 and 3.10 of the FBI Laboratory Quality Assurance Manual and with the LOM Practices for Retaining Case-Related Documentation. Appendix B contains the forms used for casework and database documentation in the unit. This is a controlled document.

10 DNA Analysis Unit II Quality Assurance Manual DNA Analysis Unit II Quality Assurance Manual 2.doc Issue Date: 03/26/08 Revision: 2 Page 10 of Case Assignment - mtdna Casework The assignment of cases, as well as the Case Acceptance Policy in the unit, is the responsibility of the Unit Chief or designee. Cases are assigned to Examiners on the basis of Examiner expertise, availability, prior related examinations, liaison conducted by DNAU II personnel, and/or other relevant factors. Case assignments are logged into the DNAU II Case Management System and distributed to Examiners by the Unit Chief or designee Case Packet Assembly - mtdna Casework Case-related administrative and examination documentation are assembled when submitting a case for technical review, administrative review and subsequent filing. Serialized correspondence is not placed into the Supporting Documentation Envelope (1-A envelope) Administrative Documentation Typically, administrative documents associated with a case are assembled as described below, in order from top to bottom, to facilitate technical and administrative reviews. These documents do not require page numbers but must bear the FBI Laboratory number. At top of packet: 1. Technical Review Sheet (may instead be placed second) 2. Administrative Review Sheet (may instead be placed first) 3. Original report (7-1) 4. Report file copy 5. Tickler copy(ies) 6. Completed 7-2a, Laboratory Statistics Sheet ( pink sheet ) This is a controlled document.

11 DNA Analysis Unit II Quality Assurance Manual DNA Analysis Unit II Quality Assurance Manual 2.doc Issue Date: 03/26/08 Revision: 2 Page 11 of 20 Below that: 7. Laboratory Work Sheet(s) (7-2) 8. ECS search slip(s) (where present) 9. Chain of Custody Log(s) (7-243a) 10 Case check-in notes (as applicable) 11. Secondary Evidence Inventory (where DNAU II secondary evidence is recorded) 12. Case Activity and Communication Log(s) (7-245) 13. Copy of incoming communication(s) 14. Invoices, subpoenas, state laboratory reports, anthropology reports, or any other miscellaneous paperwork. These items may be placed in a clasp envelope which is marked with the appropriate FBI Laboratory number(s), contents of the envelope ( Administrative Documents is an appropriate and adequate description of the contents) and the number of pages contained in the envelope. 15. Deviation Requests (if applicable) 16. Evidence Control Unit Examination Plan If an outgoing report addresses multiple submissions of evidence (multiple laboratory numbers), each additional administrative document should be organized in the manner indicated above in the order of the laboratory number listed on the report. All administrative documents should be organized together in order by laboratory number. DNAU II secondary evidence is recorded on the Secondary Evidence Inventory. The totality of administrative documents is recorded on the 1-A envelope Examination Documentation - Biologist Notes This documentation typically consists of the appropriate Biologist notes. Items 5 and 6 below are placed in an envelope(s). All envelopes are marked with the appropriate FBI Laboratory number(s), contents of the envelope and the number of pages contained in the envelope. Items 1-4 below are numbered. The Biologist notes typically consist of the following: 1. Procedure Summary sheet 2. Extraction/Quantification sheet 3. Amplification/Cycle Sequencing sheet 4. Notes/Photos sheet(s), where applicable 5. Agilent 2100 Bioanalyzer electropherograms (with coversheets) 6. Sequence Sample sheet(s) This is a controlled document.

12 DNA Analysis Unit II Quality Assurance Manual DNA Analysis Unit II Quality Assurance Manual 2.doc Issue Date: 03/26/08 Revision: 2 Page 12 of Examination Documentation - Examiner Notes This packet (or packets) contains the appropriate Examiner notes. This documentation is placed in an envelope(s). All envelopes are marked with the appropriate FBI Laboratory number(s), contents of the envelope and the number of pages contained in the envelope. The Examiner notes typically consist of the following: 1. Case summary sheet 2. Examiner Sequencher sheets 3. Confirming Examiner Sequencher sheets (where appropriate) 4. Population database search sheets (CODIS + mito Popstats) (where appropriate) A compact disc (CD), containing at a minimum the Genetic Analyzer 3130xl analysis files, Agilent 2100 Bioanalyzer files, and Sequencher files, is also part of the Examiner notes. The CD is labeled with, at a minimum, the relevant FBI Laboratory number(s), initials of the Examiner, and the date the CD is created Sending Case Documentation to File After the case documentation has been subjected to the complete review process and the report sent to the contributor, the pink sheet(s) are sent to ECU. The remaining case documentation, with the exception of the report file copy, is placed in one or more 1-A envelopes, with appropriate information completed on it, including an accounting of the totality of pages included in the 1-A envelope. The report file copy and 1-A envelope are ultimately stored in the Laboratory file room mtdna Case Reviews Sequence Confirmation At a minimum, matching samples in cases which involve samples with concordant sequences, TEDAC cases, and those samples which will be uploaded into NDIS, must be confirmed by a second qualified Examiner. Confirmation involves independently assembling sequences for the negative control, reagent blank, and evidentiary samples from the analysis data. Separate Sequencher printouts are generated and initialed by each Examiner for the sample, as well as for the reagent blank and negative control where applicable. Confirmed positive control printouts are named with the symbols of both Examiners and marked OK and initialed by both of the Examiners. Confirmed sequence range is defined as the shortest length of double-stranded sequence obtained by two Examiners (with the exception of HV1 C-stretches, where there may be short stretches of single-stranded sequence). Confirmation is documented on the Procedure Summary Sheet by Examiner name, symbols, and date confirmed, and on the Sequencher sheets by the confirming Examiner's symbols, This is a controlled document.

13 DNA Analysis Unit II Quality Assurance Manual DNA Analysis Unit II Quality Assurance Manual 2.doc Issue Date: 03/26/08 Revision: 2 Page 13 of 20 hand-written initials and the date confirmed. If confirmation cannot be achieved, the Unit Chief or designee provides the technical guidance necessary for resolution, in accordance with LOM Practices for Scientific or Technical Casework Conflict Resolution Technical Reviews Technical reviews are performed in compliance with Section of the FBI Laboratory Quality Assurance Manual and the LOM Practices for Reviewing a Report of Examination. All mtdna examinations are technically reviewed by another qualified Examiner before the examination results are reported and submitted to the Unit Chief or designee for administrative review. The Technical Review consists of a review of all administrative and examination documentation, including data, sequence confirmation and other documents which form the basis for the conclusion(s) reached. The technical reviewer documents the review by signing and dating the Technical Review Sheet, the Procedure Summary Sheet (see relevant appendices), and the report file copy Administrative Reviews Administrative reviews are performed in compliance with Section of the FBI Laboratory Quality Assurance Manual and the LOM Practices for Reviewing a Report of Examination. Every Report of Examination (with accompanying administrative and examination documentation) is administratively reviewed by the Unit Chief or designee before it is sent to the contributor. The administrative reviewer completes, signs and dates the Administrative Review Sheet (see Appendix B) and signs and dates the report file copy after completing the review of the case Recording of mtdna Laboratory Reports and mtdna Case Information All Reports of Examination generated in the Unit are uploaded into the FBI's Automated Case Support (ACS) System. Relevant case information is recorded in the DNAU II Case Management System before the report is sent to the contributor. 12 Instrument Calibration and Maintenance The LOM Practices for Instrument Calibration and Maintenance and Sections 5.5 and 5.6 of the FBI Laboratory Quality Assurance Manual provide general guidelines for instrument calibration and maintenance. Instruments and equipment in the Unit are used in accordance with these as well as with Section 2 of the Mitochondrial DNA Analysis Protocol. This is a controlled document.

14 DNA Analysis Unit II Quality Assurance Manual DNA Analysis Unit II Quality Assurance Manual 2.doc Issue Date: 03/26/08 Revision: 2 Page 14 of The following general guidelines apply to instruments and equipment used in the Unit: An inventory is kept of all equipment and instruments used in the Unit A file or notebook for each relevant instrument and item of equipment is maintained. The information retained includes any corresponding manuals, warranty information, maintenance, service, and calibration procedures and records. This information is maintained in the Unit All equipment and instrumentation is used in accordance with the manufacturer=s instructions Individual Unit employees have specific responsibilities for the maintenance, service and calibration of equipment and instruments The QA Program Manager reviews the calibration and maintenance/service documentation. Review of the documentation is noted in the appropriate calibration notebook using the initials of the reviewer and date. Additionally, the QA Program Manager directs and documents any action deemed necessary as a result of calibration checks, maintenance, or service The Mitochondrial DNA Analysis Protocol specifies the equipment and instruments subjected to calibration and/or maintenance for mtdna casework, as well as the procedures used for each. 13 Quality Control of Reagents and Supplies DNAU II complies with Sections 4.6 and of the FBI Laboratory Quality Assurance Manual and Section of the LOM Practices for the Examination of Evidence with regard to quality control of reagents and supplies Documentation of Receipt The receipt of all chemicals/supplies must be recorded in the DNAU II Chemicals and Reagents Log. All chemicals/supplies are labeled to indicate date received, date opened for use, Examiner or Biologist=s initials and storage conditions. When appropriate, QC procedures are performed, and the approval for use in casework is indicated by initialing and dating the label and notation in the log Documentation of Reagents Labeling of reagents includes the identity of the reagent, lot number, date received or prepared, preparer=s initials (where appropriate), storage conditions, start date, and expiration date (one year This is a controlled document.

15 DNA Analysis Unit II Quality Assurance Manual DNA Analysis Unit II Quality Assurance Manual 2.doc Issue Date: 03/26/08 Revision: 2 Page 15 of 20 if not otherwise specified). The start date should be recorded in the Chemicals and Reagents Log. All stock reagents will be discarded if not used within a year of the date prepared. When appropriate, QC procedures are performed, and the approval for use in casework is indicated by initialing and dating the label and notation in the log Critical Reagents Section 2 of the Mitochondrial DNA Analysis Protocol identifies the critical reagents used in mtdna analysis and describes quality control procedures to ensure suitability for casework. 14 Court Testimony Monitoring Examiners in the DNA II Unit have their courtroom testimonies evaluated in accordance with Sections 3.11 and of the FBI Laboratory Quality Assurance Manual and with the LOM Practices for Court Testimony Monitoring. An External Evaluation of Testimony Form (7-257) and/or an Internal Evaluation of Testimony Form (7-256) is completed for at least one testimony per year for each examiner. When the form is returned to the Unit Chief, the Unit Chief will review it with the Examiner and the Examiner will sign it. Testimony Monitoring forms are maintained in the Unit, with the lower portion being redacted after the Performance Appraisal Review each year. 15 Addressing Complaints Complaints regarding DNAU II services are addressed by the Unit Chief or Examiners, as appropriate, in compliance with Section 4.8 of the FBI Laboratory Quality Assurance Manual (and with the LOM Practices for Corrective Action and the LOM Practices for Preventive Action, where appropriate). They are authorized to recommend, direct, and/or implement appropriate corrective or preventive action. 16 Proficiency Testing DNAU II complies with the proficiency testing requirements in Sections and 5.9 of the FBI Laboratory Quality Assurance Manual, the LOM Practices for Open Proficiency Testing, and the FBI Standards. If an Examiner or Biologist is on extended leave, then he or she shall successfully complete a proficiency test prior to resuming casework. This is a controlled document.

16 DNA Analysis Unit II Quality Assurance Manual DNA Analysis Unit II Quality Assurance Manual 2.doc Issue Date: 03/26/08 Revision: 2 Page 16 of Scope Each Examiner and Biologist qualified to perform mtdna casework is subject to the Unit's external proficiency testing program Testing Areas and Frequency Each person subject to proficiency testing is tested in the area of mtdna analysis twice a year, at a frequency not to exceed six months from the completion of one test (defined as the provider=s due date) to the distribution of the subsequent test. All tests are external and supplied by an American Society of Crime Laboratory Directors/Laboratory Accreditation Board (ASCLD/LAB)-approved provider. An internal proficiency test may be used when a person returns from extended leave to fulfill the requirement stated above. Each person tested is tested to the full extent to which she or he performs casework. Typical test materials include blood samples, saliva samples, and/or hairs. Because the Unit does not analyze samples presumed to contain DNA mixtures, such as vaginal swabs, proficiency testing is not conducted on such samples Test Evaluation Proficiency tests are evaluated by the QA Program Manager using the manufacturer's results and according to the criteria set forth in the FBI Standards, and records are maintained in accordance with these standards as well as with the LOM Practices for Open Proficiency Testing. Any tests taken by the QA Program Manager are evaluated by the Unit Chief. The FBI Laboratory Proficiency Test Program Manager also evaluates external proficiency tests according to the LOM Practices for Open Proficiency Testing. The evaluation and compliance with record maintenance requirements are documented on the DNAU II Proficiency Test Evaluation form (Appendix C). 17 Audits 17.1 The audit process is designed to ensure that the quality assurance objectives of DNAU II are met through a critical review of personnel qualifications, the work conducted by the Unit staff, and the laboratory facility. This is accomplished primarily by review of Unit documentation The Unit is audited annually in accordance with the criteria set forth in the FBI Standards, including the provision of audits performed by an external agency every other year. Documentation maintained by DNAU II includes the audit checklist, audit report, and responses (including any corrective action taken) to findings raised in the audit report Audits of DNAU II performed as part of an ASCLD/LAB accreditation inspection meet the external audit requirement of the FBI Standards. This is a controlled document.

17 DNA Analysis Unit II Quality Assurance Manual DNA Analysis Unit II Quality Assurance Manual 2.doc Issue Date: 03/26/08 Revision: 2 Page 17 of In addition to audits conducted to comply with the FBI Standards, DNAU II is subjected to periodic audits of the quality system conducted by the Quality Assurance and Training Unit, in accordance with Section 4.14 of the FBI Laboratory Quality Assurance Manual and the LOM Practices for Internal Audits. Documentation associated with these audits is maintained by the Unit and the Quality Assurance and Training Unit Under the direction of the DNAU II Unit Chief, the QA Program Manager is responsible for addressing all suggestions, recommendations, findings and possible corrective actions as a result of the audit process. The Unit Chief is responsible for ensuring that appropriate corrective action is taken when necessary and documented. 18 Corrective Action DNAU II acknowledges, documents, and resolves any technical problem involving casework or proficiency testing in accordance with Section 4.11 of the FBI Laboratory Quality Assurance Manual and the LOM Practices for Corrective Action and Practices for Open Proficiency Testing. 19 Measurement Uncertainty Measurement uncertainty does not apply to examinations conducted in DNAU II. 20 DNAU II Examination Notes Abbreviations The following abbreviations may be encountered in DNAU II examination documentation: A - adenine abna - analyzable but not alignable am - ante meridiem, the period from 12 midnight to 12 noon amp amplify or amplification bp - base pair(s) C - cytosine CE - capillary electrophoresis CODIS + Mito - Combined DNA Index System for mitochondrial DNA (refer to definition of DNA) cm - centimeter db - database dil - dilution CS - cycle sequence(d) DNA - deoxyribonucleic acid This is a controlled document.

18 DNA Analysis Unit II Quality Assurance Manual DNA Analysis Unit II Quality Assurance Manual 2.doc Issue Date: 03/26/08 Revision: 2 Page 18 of 20 DNAU II/DNAU 2 - DNA Analysis Unit 2 (refer to definition of DNA) FBI - Federal Bureau of Investigation G - guanine HV1(HVI), HV2(II), HV1a, HV1b, HV2a, HV2b - Hypervariable regions 1, 2, 1a, 1b, 2a, and 2b I2 - the second injection for a particular evidence extract (I3 and so on may be used as needed) K - evidentiary item of known origin L - liter λ microliter mm - millimeter mtdna - mitochondrial DNA (refer to definition of DNA) na - not analyzable or not applicable nc - negative control ng - nanogram(s) nmol nanomole or nanomoles NMPDD - National Missing Persons DNA Database (refer to definition of DNA) np - nucleotide pair(s) P2 - the second amplification for a particular evidence extract (P3 and so on may be used as needed) p - page pc - positive control PCR - polymerase chain reaction pg - picogram or page(s) pm - post meridiem, the period from 12 noon to 12 midnight Q - evidentiary item of questioned origin quant quantify or quantification rb - reagent blank rcrs - revised Cambridge Reference Sequence rec=d - received ref - reference ri - re-injection rr - re-run S2 - the second cycle sequencing reaction for a particular evidence extract (S3 and so on may be used as needed) SEI secondary evidence inventory seq - sequence or sequenced std - standard T - thymine ul microliter WCR whole control region X2 - the second extraction for a particular item of evidence (X3 and so on may be used as needed) This is a controlled document.

19 DNA Analysis Unit II Quality Assurance Manual DNA Analysis Unit II Quality Assurance Manual 2.doc Issue Date: 03/26/08 Revision: 2 Page 19 of 20 Rev. # Issue Date History 0 06/09/06 Original document issued. 1 11/12/07 Editorial changes throughout document. Updated mission statement in Section 1.1. Deleted Information Technology and Health and Safety as unit programs and added Regional Mitochondrial DNA Laboratories and Mass Spectrometry programs in Section 2.2. Updated Section 4.3 to require Unit Chief to assess training needs. Addition of limited facilities services employees with access to evidence storage areas as necessary for maintenance in Section 5.1. Updated temperature information in Section 8.2. Removed references to Quantiblot procedure from Sections 8.3 and as it is no longer in use. Removed instructions in Section 8.6 for return of evidence submitted prior to 03/11/03, since the unit no longer has any evidence received prior to this date. Updated Section to include additional documentation to be included in Administrative documentation packet. Modified proficiency test requirements in Section 16.2 for an Examiner or Biologist on extended leave. Updated Biologist workbook to remove sections for Quantiblot procedure as it is no longer in use. Added calculation sheets for new procedure and revision numbers to each page of workbook. 2 03/26/08 Added mtdna Operations to list of programs in the Unit in Section 2.2. Updated Section to reflect new room location for Telekey box. Updated Section to reflect new room location for temporary storage of secondary evidence. Added requirement for Technical Reviewer to document review by also signing report file copy in Section Addition of commonly used abbreviations to Section 20. This is a controlled document.

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