EZChrom Edition with OpenLAB ECM

Transcription

1 OpenLAB CDS Integration REVOLUTIONIZE of Agilent YOUR DESKTOP OpenLAB CDS SEQUENCING WORKFLOW EZChrom Edition with OpenLAB ECM HALOPLEX TARGET Compliance ENRICHMENT with SYSTEM 2 CFR Part What a Difference a Day Makes Complete Target Enrichment in Less Than 6 hours Technical Note Introduction Highlights Expanded capture size enrich thousands of targets, all in a single tube Intuitive Design Wizard create panels in less than minutes Compatibility with desktop sequencers and high-throughput NGS platforms Scalable protocols 96-samples ready for sequencing in less than a day, with Agilent automation solutions Performance and reliability greater specificity, uniformity, and variant calling Immediate implementation no dedicated capital equipment required Part in Title 2 of the Code of Federal Regulations includes the US Federal guidelines for storing and protecting electronic records and applying electronic signatures. The intent of these Summary guidelines is to ensure that applicable electronic records are reliable, authentic and maintained with high integrity. The Agilent HaloPlex Target Enrichment System enables fast, simple, and efficient This analysis technical of genomic note describes regions features of interest and functionality for a large of number Agilent OpenLAB of samples, CDS covering EZChrom Edition, revision A.4.2 or higher, in combination with Agilent OpenLAB Enterprise Content thousands of exons per sample. It uses single-tube target amplification and removes Manager (ECM) revision..2 SP HF or higher, which enable them to meet the guidelines the need for library preparation to reduce total sample processing time and cost of 2 CFR Part. without the need for dedicated instrumentation. HaloPlex is compatible with This different document desktop examines sequencing each section and high-throughput of 2 CFR Part and platforms. provides It a also recommended expands your remediation approach using the integration of OpenLAB CDS EZChrom Edition (revision options, with the availability of 96 indexes and an automation protocol, to deliver the A.4.2 or higher) with Agilent OpenLAB ECM (revision..2 SP HF or higher). flexibility you need to efficiently run your experiments. Finally, you can access an Agilent assay with OpenLAB premium CDS EZChrom performance, Edition high revision specificity, A.4.2 provides uniformity, a tight and and variant seamless detection. integration into Agilent OpenLAB ECM. This integrated solution provides the necessary controls PREMIUM for PERFORMANCE managing system access, user management, data transfer handling and audit trail functionality, while Agilent OpenLAB ECM ensures secure record keeping and data archival. Design Size Range Up to 5Kb (>2,5 exons);.5mb coming soon Specificity (% bases on-target) > 99 %* powerful Uniformity search (coverage capabilities, at % of data mean) viewers and archiving > 9 %* features for all data in the lab independent covered from at its X data format. OpenLAB ECM > 98 is proven %* to satisfy compliance needs as mandated covered by regulations X such as 2 CFR Part. > 97 It has %* been implemented by many leading life % science Bases covered companies at 2X for this reason. > 96 %* *Performance data was collected for 2 different designs, with sizes ranging from 4.4kb to 4kb. Agilent OpenLAB ECM is an electronic library. It provides a secure compliant data repository, While the integration of OpenLAB CDS EZChrom Edition with OpenLAB ECM provides significant % and extensive compliance tools to facilitate compliance, the ultimate responsibility rests 8% with the enterprise employing the technology. Complete compliance therefore requires that 6% the enterprise employing this functionality have the appropriate internal Standard Operating Procedures (SOP). Articles that can only be satisfied by an SOP are indicated as 4% such in the detail below. Table gives an overview of the different sections of the Part- 2% regulations, and which of them are applicable for OpenLAB CDS EZChrom Edition with % OpenLAB ECM. Covered by Design on Target Covered x Covered 2x 4.4 Kb 2 Kb 6 Kb 2 Kb 28 Kb 96 Kb Covered 2x Covered at % of Mean Covered at 2% of Mean

2 Identification of Unknown Organic Pollutants A key strength of the method described here is its ability to simultaneously identify 2 unexpected CFR Part organic sections pollutants. As addressed demonstrated by in the Figure integration, an unexpected of Agilent OpenLAB CDS EZChrom Edition 2.5 (revision contaminant, A.4.2 metoprolol, or was higher) identified with by accurate Agilent mass OpenLAB measurement ECM and (revision isotope..2 SP HF or higher) 2 pattern matching. The identity of metoprolol is further confirmed by matching its MS/MS spectrum to the Broecker, Herre & Pragst Forensics/Toxicology Personal.5 Compound Database and Library (Figure 4). The integration of OpenLAB CDS EZChrom Edition with OpenLAB Electronic Electronic Possible scenarios with record only signature based ECM enables the operation of OpenLAB CDS EZChrom Edition in full x EZChrom 2 +ESI Product operated Ion (2.752, in a min, 2 Scans) (without Frag=225.V [email protected] upon User (268.98[z=] ID -> **) closed system support of all compliance requirements mandated by 2 CFR Part signature) and Password.75 for a closed system. In particular it ensures: ,.2, Counts vs. Acquisition time (min) (Scope, implementation, definition) Accurate and complete copies of records Figure 2. Extracted ion chromatogram of x 2 +ESI Product Ion (2.752, min, 2 Scans) Frag=225.V [email protected] (268.98[z=] -> **) phenazone at the ng/l level, with the (Controls for closed systems) Versioning of all relevant records for traceability structure of the compound (inset). -. Preservation of records between their creation and their automatic (Controls - for open systems) transfer to OpenLAB ECM immediately after acquisition, x 2 Metoprolol C5H25NO + Product Ion Frag=5.V CID@2. ForensicsTox_AM_PCDL.cdb reprocessing or interactive modifications x 4 Metoprolol: +ESI Scan.5.75 (Signature manifestations) Controlled copies of the data (Signature record linking) Mandatory login to OpenLAB ECM before allowing appropriate.8 access to EZChrom..7 (e-sig general requirements).6 Figure 4. Identifi cation of metoprolol can be further confi rmed by MS/MS and retention time Records of changes captured in user-independent time-stamped matching to a reference compound (top: sample spectrum, middle: mirror spectrum, bottom:.2 (a).4 library spectrum). audit trails (e-sig not biometric).2 (b) These settings can be configured during or after installation to meet Broad (e-sig biometric) Applicability specific standard operation procedures and security guidelines. The. 655 (a), Q-TOF (b), (d) with Jet Stream and ifunnel technology delivers the This utmost includes customizable EZChrom security Counts vs. roles Mass-to-Charge and privileges (m/z) (Controls for ID codes / passwords) sensitivity for a broad range of target compounds (Table ), enabling ultra-trace that provide restricted access Figure. to EIC EZChrom of an unexpected functionality contaminant, to individual metoprolol, with a signal-to-noise ratio of 46:. compound. (c), (e) identification and quantitation. users or groups. Changes Red to squares the security represent configuration the theoretical can be isotope made (Token cards & other ID devices) only by a dedicated system intensity administrator. and position. Details and recommendations for configuration of the system are outlined in the manual Table : Table. LOD and LOQ values (ng/l) for a broad range of target compounds. Applicable sections of 2 CFR Part for OpenLAB CDS EZChrom Edition/OpenLAB ECM Compound operated Name in a closed system ( = LOD applicable, LOQ = not Compound applicable) OpenLAB CDS Administration Guide, Part Number M85-9, Name LOD LOQ available on the OpenLAB CDS EZChrom Edition installation media. Atenolol Sulfamethoxazole Sotalol Carbamazepine, epoxide The Metronidazole integration of OpenLAB CDS.7EZChrom.9 Edition Phenacetin with OpenLAB ECM was not.8specifically.5 designed for operation in an open system. Ronidazole Bisoprolol NOTE: In this technical note the term EZChrom (in the context of integration with OpenLAB ECM) always refers to OpenLAB CDS EZChrom Sulfadiazine Propranolol Edition with the feature set for the integration with OpenLAB ECM enabled. Trimethoprim.2.2 Carbamazepine 2.9 Sulfamerazine Betaxolol Sulfadimidine n,n Diethyl--methylbenzamide Phenazone (DEET) Pentoxifylline Bezafibrate Metoprolol Crotamitron For Research Use Only. Not for use in diagnostic procedures. Furazolidone Diazepam.4.97 Dapson Fenofibrate.5 2. *Calculated based on the DIN 2645 using a program courtesy of Dr. Joachim Kleiner. ( x Phenazone: +ESI EIC(89.22) SNR (4.8min) = Agilent Technologies, Inc. 22 Published in the USA, April 5, EN

3 Identification. Controls of for Unknown Closed Systems Organic Pollutants Phenazone: +ESI EIC(89.22) x 4 SNR (4.8min) = 68. A Section key strength Question of the method described here OpenLAB is its ability CDS EZChrom to simultaneously Edition/OpenLAB identify ECM Integration 4.8 unexpected organic pollutants. As demonstrated in Figure, an unexpected Yes. Agilent Technologies has extensively validated the performance of EZChrom 2.5 / OpenLAB ECM with tests written to specifically contaminant, Has metoprolol, the system been validated was identified in order by accurate evaluate accuracy, mass reliability measurement and consistent performance. and isotope The integrated solution of EZChrom with OpenLAB ECM incorporates the to ensure accuracy, reliability, consistent.(a) use of byte- order dependent check sums each file transfer operation to ensure 2 that records are valid and unaltered. pattern matching. intended performance, The identity and the of ability metoprolol is further confirmed by matching its to discern invalid or altered records? Agilent Technologies recommends making use of Installation Qualification and Operation Qualification (IQ/OQ) service for EZChrom / MS/MS spectrum to the Broecker, Herre & OpenLAB Pragst ECM Forensics/Toxicology to validate the on-site system per Personal compliance requirements..5 Compound Database and Library (Figure 4). Is the system capable of generating Yes. OpenLAB ECM stores all data types, from raw machine data to printable reports. All files are stored complete and unaltered in accurate and complete copies of all the original format. x.(b) 2 +ESI Product required Ion records (2.752, in both human min, 2 readable Scans) Frag=225.V [email protected] (268.98[z=] -> **) Raw, meta and result data generated by EZChrom are stored and managed in OpenLAB ECM. Printed reports can be stored as and electronic form suitable for inspection, PDF files which can be made available for review without the source application being installed on the client machine. Depending.75 review and copying by the FDA? on the data type, a wide selection of file viewers is available to view the original electronic record without the original application Yes. Raw data, 9.78 meta data and result data generated by EZChrom are stored and managed in OpenLAB Counts ECM. vs. Acquisition Data stored in time OpenLAB (min) ECM resides in a protected storage location or archive media or both. When archived, the media may be on-line, near-line or off-line. Are the records protected for accurate Regardless of the physical location of the data, it remains searchable to all users Figure with appropriate 2. Extracted privileges. ion chromatogram The individual users do of not x 2 +ESI Product Ion (2.752, min, 2 Scans) Frag=225.V [email protected] (268.98[z=] -> **).(c) and convenient retrieval throughout the have access to the physical storage location of the files. phenazone at the ng/l level, with the record retention period? The functionality 9.78 is complemented by additional procedural controls that should structure be defined of and the implemented compound by the (inset). system administrator based on company-wide security policies. These policies should manage practices such as access to client computers and password - renewal frequency. - Yes. Each user is identified by a unique user ID and password combination. Access to EZChrom / OpenLAB ECM requires the entry x Is system access limited to authorized.(d) 2 Metoprolol C5H25NO + Product Ion Frag=5.V CID@2. ForensicsTox_AM_PCDL.cdb of both identification components: user ID and password. In EZChrom / OpenLAB ECM, all file and software functionality access is individuals? x 4 Metoprolol: +ESI Scan controlled by privileges and roles assigned to individual users or groups of users. The system administrator determines levels of access Is there a secure, computer-generated Yes. All activities 9.67 in EZChrom / OpenLAB ECM are recorded in secure, computer-generated, time-stamped audit trails. Audit trails.25 audit trail that independently records the date exist 76.7 for system, instruments, methods, sequences, data and report templates. The data audit trail is embedded within the data itself.(e) and time of operator entries and actions that to ensure long-term retention and association. Entries in the EZChrom / OpenLAB.9 ECM audit trails are non-editable, non-deletable. 6 7 create, 8 modify, 9 or delete electronic 2 records? Even 7 the 8removal 9 2 of records 2 22 from 2 OpenLAB ECM 26 by an 27 authorized user does.8 not affect existing entries in the audit trail..7 When records are changed, is previously Yes. All entries in the EZChrom / OpenLAB ECM audit trails are non-editable, non-deletable. Even the removal of records from.(e).6 Figure 4. Identifi recorded cation information of metoprolol left unchanged? can be further confi OpenLAB rmed ECM by by MS/MS an authorized and user retention does not affect time existing entries in the audit trail. matching to a reference compound (top: sample spectrum, middle: mirror spectrum, bottom: Are electronic audit trails saved at least as Yes. Audit trail entries are stored in the OpenLAB ECM repository as part of a.4 file s metadata and are kept throughout the electronic library.(e) spectrum) long as their subject electronic records and records retention period. The audit trail may be reviewed and printed. The OpenLAB. ECM audit trails are unbreakably linked to the available for agency review and copying? record. System-related activities such as logon events are unbreakably linked.2 to the system Are operational system checks used to Yes. In all EZChrom / OpenLAB ECM functions, when a sequencing of events is required, system checks enforce it. In EZChrom users Broad.(f) Applicability enforce permitted sequencing of steps are stepped through sequences and events; data cannot be acquired unless parameters 268are within 268.5valid 269 instrument and integration and events? ranges. Users are prompted with an error message when steps are performed out of sequence. The 655 Q-TOF with Jet Stream and ifunnel technology delivers the utmost Are authority checks in place to ensure Figure. EIC of an unexpected contaminant, sensitivity for that a only broad authorized range individuals of target can use compounds Yes. Users (Table cannot gain ), access enabling to the software ultra-trace modules of EZChrom / OpenLAB metoprolol, ECM without with a valid a signal-to-noise user ID, password and ratio account. of 46:. the system, electronically sign a record, compound.(g) identification access the operation and or computer quantitation. system input or output device, alter a record, or perform the current operation? Table. LOD and LOQ values (ng/l) for a broad range of target compounds. Once logged in, that user s access to files and software functionality (including Red but squares not limited represent to signing a the file, theoretical inputting values, isotope or altering a record) is determined by the privileges assigned to the user. intensity and position. Are device checks used to determine, as.(h) Compound Name appropriate, the validity LOD of the source LOQ of data Yes. The integration with instruments through comprehensive automation and control provides various levels of device and validity checks depending on the instrument make and model. Compound Name LOD LOQ Atenolol or operational instruction? User entry fields provide feedback to the user about the entry types and ranges that are valid for that field in EZChrom. Sulfamethoxazole Sotalol Carbamazepine Yes. Records of the educational, epoxide and employment.82 history.52of Agilent Technologies employees are verified and kept with personnel Do the persons who develop, maintain, records that can be made available during an on-site audit. In addition, all Agilent Technologies employees who work with regulations Metronidazoleor use electronic records.7 and signature.9 Phenacetin have attended training workshops for regulatory.8 requirements..5.(i) systems have the education, training, Ronidazole and experience to perform 2.65 their assigned 9.7 Bisoprolol Users of EZChrom / OpenLAB ECM at a customer.8 location 5.52 will be required to show records or education, training and/or experience tasks? with the system. Agilent provides a basic familiarization during the installation of the product for system users. Training courses for Sulfadiazine Propranolol administrators as well as users are available Trimethoprim.2.2 Have written policies that hold individuals Carbamazepine 2.9 Sulfamerazineaccountable and responsible 2.55for actions 9.9 Betaxolol. It is the responsibility of the organization.6implementing 4.4 electronic signatures to develop written policies which ensure that.(j) initiated under their e-signatures in order Sulfadimidine individuals responsible for signing documents understand that their electronic signature is as equally binding as their handwritten signature. to deter record and signature 2.59falsification 9.5 n,n Diethyl--methylbenzamide been established and followed? Phenazone (DEET) PentoxifyllineAre there adequate controls.7over the 4.47 Bezafibrate distribution of, access to, and use of. While documentation is available for the integration of EZChrom / OpenLAB ECM for users and administrators, controls over the. Metoprolol (k)() documentation for system 6 operation 2.5 Crotamitron and storage and distribution of this material are the responsibility of the organization that implements and uses the system. maintenance? For Research Use Only. Not for use in diagnostic procedures. Furazolidone Diazepam.4.97 Dapson Are there formal revision.85 and change 4.7 control Fenofibrate.5 2. Yes. Agilent Technologies quality process includes formal written revision and change control procedures for system documentation. procedures to maintain an audit trail that Agilent Technologies, Inc. 22. (k)(2) The process includes the use of OpenLAB ECM for development of system documentation. All revisions to the documents kept are time *Calculated based documents on the DIN 2645 time-sequenced using a program development courtesy and of Dr. Joachim Kleiner. ( Published in the USA, April 5, 22 stamped and audit-trailed. modification of systems documentation? EN

4 Identification. Controls of for Unknown Open Systems Organic Pollutants Phenazone: +ESI EIC(89.22) Section Question OpenLAB CDS EZChrom Edition / OpenLAB ECM integration x 4 SNR (4.8min) = 68. A key strength of the method described here is its ability to simultaneously identify 4.8 Are there procedures and controls used Yes. When a file is transferred to or within OpenLAB ECM, a byte-order dependent checksum is calculated on the file in its source unexpected to organic protect the pollutants. authenticity, integrity As and demonstrated in Figure, an unexpected location. A copy of the file is made of the file in the destination location where 2.5 a second checksum is calculated. The two values contaminant,. confidentiality metoprolol, of the electronic was identified records by accurate are compared mass and only if measurement they are identical, is the and transfer isotope complete. If the values do not match, an error message is generated. from their creation point to the point of their receipt? The integration of EZChrom / OpenLAB ECM was not specifically designed for 2 pattern matching. The identity of metoprolol is further confirmed by matching its operation in an open system. MS/MS spectrum to the Broecker, Herre & Pragst Forensics/Toxicology Personal.5 Yes. OpenLAB ECM supports the use of Secure Socket Layer (SSL) encryption for security during data transmission. SSL breaks a Compound Database Are additional measures and Library used to ensure (Figure the 4). single file into very small data packets. These data packets are individually encrypted with configurable 64-bit or 28-bit encryption confidentiality of the electronic records from. before being transmitted. On the receiving side the data packets are decrypted and reassembled. The integration of EZChrom with the point of their creation to the point of their OpenLAB ECM supports SSL encryption. x 2 +ESI Product receipt? Ion (2.752, min, 2 Scans) Frag=225.V [email protected] (268.98[z=] -> **) The integration of EZChrom with OpenLAB ECM was not specifically designed for operation in an open system Signature Manifestation 9.78 Counts vs. Acquisition time (min) Section Question OpenLAB CDS EZChrom Edition / OpenLAB ECM integration Figure 2. Extracted ion chromatogram of x 2 +ESI Product Ion (2.752, min, 2 Scans) Frag=225.V [email protected] (268.98[z=] -> **) Yes. The EZChrom electronic signature manifestation includes: phenazone at the ng/l level, with the The user 9.78 ID in addition to the full name of the signer structure of the compound (inset). The date and time that the signature was applied - Do the signed electronic records contain The user configurable meaning associated with the signature - information associated with the signing that clearly indicates the following: An esignature time-out is provided for added security. x 2 Metoprolol C5H25NO + Product Ion Frag=5.V CID@2. ForensicsTox_AM_PCDL.cdb.5(a) printed name of signer In addition, EZChrom result sets or SSIZip files can be electronically signed in x OpenLAB 4 Metoprolol: ECM. The +ESI OpenLAB Scan ECM electronic signature manifestation includes: date and time the signature was executed The user ID in addition to the full name of the signer meaning associated with the signature? The signer s 9.67 title The date and time that the signature was applied The location 8 9 where 2 the 2 signing 22 2 occurred The user configurable meaning associated with the signature.7 Yes. Both the EZChrom machine-readable data and the human readable report.6contain the name, date and meaning of the EZChrom Figure 4. Identifi cation of metoprolol can be further confi Are these items part of any human readable electronic rmed signature. by MS/MS and retention time matching.5(b) to a form reference of the electronic compound record? (top: sample spectrum, middle: mirror spectrum, bottom: In addition, the optional esignature Plug-in for Adobe Acrobat places a visible.4 signature manifestation on all human readable forms of library spectrum). the document, electronic display and printed form.. Broad.7 Signature/Record Applicability Linking Section Question OpenLAB CDS EZChrom Edition / OpenLAB ECM integration The 655 Q-TOF with Jet Stream and ifunnel technology delivers the utmost Yes. Signed records have a unique checksum that prevents signatures from being excised, copied or otherwise transferred. OpenLAB Is the electronic signature linked to its Figure. EIC of an unexpected contaminant, sensitivity for a broad range of target compounds ECM will not (Table recognize ), a signature enabling that was ultra-trace applied outside its own electronic signature plug-ins. respective electronic record to ensure that metoprolol, with a signal-to-noise ratio of 46:. compound.7 identification the signature cannot and be excised, quantitation. copied or The EZChrom electronic signature is stored in the data file and is unbreakably Red linked squares to the EZChrom represent data even the outside theoretical of OpenLAB isotope ECM. The optional esignature Plug-in for Adobe Acrobat encrypts the signature within intensity the document and position. to prevent the signature from being otherwise transferred to falsify an electronic record by ordinary means? excised or copied to another document. Table. LOD and LOQ values (ng/l) for a broad range of target compounds.. Compound Electronic Name Signatures: LODGeneral LOQ Requirements Compound Name LOD LOQ Section Atenolol Question Sulfamethoxazole OpenLAB CDS EZChrom Edition 2.97 / OpenLAB.84 ECM integration Sotalol Carbamazepine Yes. EZChrom as well, as OpenLAB epoxide ECM uses.82the user.52 ID / password combination unique to each user in the electronic signature Is each electronic signature unique to one feature. User names in EZChrom / OpenLAB ECM are required to be unique and cannot be reused or reassigned to another individual.. Metronidazole (a) individual and not reused.7 by, or reassigned.9 to, Phenacetin.8.5 anyone else? Whether EZChrom / OpenLAB ECM uses the company s Windows NT logins to validate users or OpenLAB ECM administrated users, Ronidazole Bisoprolol no two users can have the same user ID /.8 password 5.52 combination. Sulfadiazine Propranolol Are the identities of the individuals verified Trimethoprim prior to the establishment,.2 assignment,.2 and Carbamazepine. This is the responsibility of the organization that 2.9 plans, implements and operates the system. Such a verification process. (b) certification of an individual s electronic is a system requirement that is set before implementing electronic signature procedures or assigning electronic signature Sulfamerazinesignature or any element 2.55 of an electronic 9.9 Betaxolol privileges to an individual Sulfadimidinesignature? n,n Diethyl--methylbenzamide Phenazone Has the company delivered.47 its corporate 2.4 (DEET). (c) electronic signature certification letter to the FDA? Pentoxifylline Bezafibrate Metoprolol (c) Is it in paper form with a 6 traditional 2.5 handwritten Crotamitron It is the company s responsibility, before submitting electronically signed documentation to the FDA, to register their intent (2) signature? to use electronic signatures. In addition, training programs must be in place For Research to ensure Use that users Only. signing Not for documents use in diagnostic electronically procedures. Furazolidone Diazepam.4.97 understand the legal significance of their electronic signature. Can additional certification or testimony be. Dapson(c) provided so that a specific.85 electronic 4.7 Fenofibrate.5 2. signature (2) is the legally binding equivalent of the signer s *Calculated based on the DIN 2645 using a program courtesy of Dr. Joachim Kleiner. ( handwritten signature? Agilent Technologies, Inc. 22 Published in the USA, April 5, EN

5 How It Works Step. Four.2 simple Electronic steps with Signature no library Components prep (Figure and ): Controls. Digest sample DNA Section Question OpenLAB CDS EZChrom Edition/OpenLAB ECM Integration A DNA sample is fragmented using restriction enzymes, and denatured. Does the e-signature employ at least two.2 (a) Yes. The EZChrom as well as the OpenLAB ECM electronic signature tools distinct identification components such as Step require 2. that the user enters two distinct identification 2. (i) Hybridize probes components; unique user ID and password. user ID and password? The probe library is added and hybridized to the targeted fragments. Each probeis an oligonucleotide When an individual designed executes a to series hybridize of to both ends of a targeted DNA restric tion signings during a single, continuous period of.2 fragment, (a) Yes. When an individual signs the first of a series of documents during a single period of controlled access the user is required to controlled thereby system guiding access, is the first targeted signing fragments to form circular DNA molecules. ()(i) enter both signature components: user ID and password. The probe executed also contains using all the electronic a method-specific signature sequencing motif that is incorporated components? during cir cularization. In addition, a sample barcode sequence is incorporated in Step. this step. When an individual executes a series of signings during a single, continuous period of Yes. When an EZChrom / OpenLAB ECM user executes a series of continuous electronic signatures, which are defined as signatures.2 (a) controlled system access, is each subsequent. executed within a period of time determined by the system administrator, they are required to enter user ID, password and reason on the ()(i) Purify and signing ligate executed targets using at least one electronic first signature only. Each subsequent signature requires only the user s password, which is known only to the user. signature component that is only executable The HaloPlex by, and probes designed to are be used biotinylated by, the individual? and the targeted fragments can therefore be retrieved with magnetic streptavidin beads. The circular molecules are then closed When an individual executes a series of Step 4. by ligation, signings a very not performed precise during reaction a single, that ensures Yes. When that an EZChrom only / OpenLAB perfectly ECM hybridized user executes a series of non-continuous electronic signatures, which are defined as.2 (a) continuous period of controlled system signatures executed outside of a period of time determined by the system administrator, they are required to enter user ID, ()(ii) fragments are circularized. access, does each signing executed require password and reason on each signature. all signature components? 4. Amplify targeted fragments with PCR Yes. OpenLAB ECM can be configured such that an administrator can assign an initial password to a user for a new account or Only circular Are controls DNA in targets place to ensure are that amplified, only providing forgotten password, an enriched but the user is and required barcoded to change that password on their first login. In this way the user ID / password.2 (a) their genuine owners can use the electronic combination is known only to the individual. (2) amplification signature? product that is ready for sequencing. Whether OpenLAB ECM uses the company s Windows NT logins to validate users or OpenLAB ECM administrated users, no two users can have the same user ID / password combination. Figure. HaloPlex workflow. HaloPlex Delivers Superior and Uniform Coverage Are the electronic signatures to be Yes. EZChrom / OpenLAB ECM uses the user s user ID and password to initiate the electronic signature. An EZChrom / OpenLAB administered and executed to ensure that the ECM user s password is stored encrypted within the database and is displayed as asterisks in all locations within the software..2 (a) attempted use of an individual s electronic A. () signature by anyone other than its genuine OpenLAB ECM can be configured B. such that an administrator can assign an initial password to a user for a new account or forgotten owner requires the collaboration of two or password, but the user is required to change that password on their first login. In this way the user ID / password combination is more individuals? known only to the individual..2 (b) Are electronic signatures based on biometrics designed to ensure that only their genuine owners can use them?. Neither EZChrom nor OpenLAB ECM support signatures based on biometrics at this time.. Controls for Identification Codes/Passwords Section Question OpenLAB CDS EZChrom Edition/OpenLAB ECM Integration Are controls in place to ensure the uniqueness of each combined identification code and Yes. EZChrom / OpenLAB ECM requires users to authenticate with user ID and password.. (a) password maintained, such that no two Whether EZChrom / OpenLAB ECM uses the company s Windows NT logins to validate users or OpenLAB ECM administrated individuals have the same combination of users, no two users can have the same user ID / password combination. identification code and password? Figure 2. HaloPlex delivers high specificity, uniformity, and sequencing coverage for any design size, and across all target regions. (A) Enrichment of all exons within the CFTR gene, for 5 different samples. (B) Contiguous and highly repetitive region (~5kb) enriched with HaloPlex. Yes. If using Windows domain authentication, password renewal interval is configured as part of the Windows password policy setup. The administrator can define a time frame in which passwords are periodically revised automatically. Users are prevented from reusing passwords.. (b). (c) Are controls in place to ensure that the identification code and password issuance is periodically checked, recalled, and revised? Are there loss management procedures in place to electronically disable lost, stolen, missing, or otherwise potentially compromised tokens, cards, and other devices that bear or generate identification code or password information? If users are administrated in OpenLAB ECM, OpenLAB ECM can be configured such that user passwords are automatically, periodically revised. Neither OpenLAB ECM nor EZChrom support devices that bear or generate identification codes, such as tokens or cards, at this time. For Research Use Only. Not for use in diagnostic procedures. Agilent Technologies, Inc. 22 Published in the USA, April 5, EN

6 How It Works Step. Four simple Are steps transaction with safeguards no library in place prep to (Figure Yes. ): EZChrom / OpenLAB ECM can be configured such that only the user knows their user ID and password. Passwords are. (d) prevent unauthorized use of passwords always displayed as asterisks and are stored encrypted within the database so that even an administrator cannot see them.. Digest sample and/or identification DNA codes? A DNA sample is fragmented using restriction Yes. OpenLAB enzymes, ECM can be and configured denatured. such that a configurable number of unauthorized access attempts lock out the user account Are transaction safeguards in place to detect and send an notification to a system administrator. and report in an immediate and urgent The Windows security policy can be configured so that a user defined number Step of Hybridize probes unauthorized access attempts locks out the user. (d) manner, any attempts at their unauthorized account and sends notification to a system administrator. The system audit trail documents general events such as logon use to the system security unit and, as The probe library is added and hybridized attempts to the to targeted the computer fragments. as well as application Each or user probeis changes, in the Windows Event log as a central audit repository for all appropriate, to organizational management? security information. This includes the system and computer ID along with the operator name and application identification, an oligonucleotide designed to hybridize to allowing both for ends an immediate of a check targeted of the potential DNA security restric leak. tion fragment, thereby guiding the targeted fragments to form circular DNA molecules. Are there controls in place to initially test The probe devices also that contains bear or generate a method-specific identification sequencing motif that is incorporated. Neither EZChrom nor OpenLAB ECM support devices that bear or generate identification codes, such as tokens or cards,. (e) code or password information to ensure that during cir at this time. they cularization. function properly In and addition, have not been a sample barcode sequence is incorporated in Step. this step. altered in an unauthorized manner?. Purify and ligate targets The HaloPlex probes are biotinylated and the targeted fragments can therefore be retrieved with magnetic streptavidin beads. The circular molecules are then closed by ligation, a very precise reaction that ensures that only perfectly hybridized fragments are circularized. 4. Amplify targeted fragments with PCR Only circular DNA targets are amplified, providing an enriched and barcoded amplification product that is ready for sequencing. Step 4. Figure. HaloPlex workflow. HaloPlex Delivers Superior and Uniform Coverage A. B. Figure 2. HaloPlex delivers high specificity, uniformity, and sequencing coverage for any design size, and across all target regions. (A) Enrichment of all exons within the CFTR gene, for 5 different samples. (B) Contiguous and highly repetitive region (~5kb) enriched with HaloPlex. For Research This information Use Only. is Not subject for use to in change diagnostic without procedures. notice. Agilent Technologies, Inc., 22 Published Agilent in the USA, Technologies, October Inc. 5, Published in the USA, EN April 5, EN