Archived. Gloves should be changed frequently during the analysis.

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1 Introduction Gloves and laboratory coats Small tools Specific clean-up and housekeeping procedures are used to help protect evidence samples from conditions and agents that might serve to destroy, deteriorate, or otherwise change the DNA to be evaluated. Gloves should be changed frequently during the analysis. Laboratory coats should be changed on a regular basis. All used lab coats are placed in designated bags in the laboratory storage area for pick-up and laundering by an outside laundry service. White lab coats are designated for use in the extraction and set up areas. Blue lab coats are designated for use in the amplification/typing room. All small tools (for example, forceps and scissors) are cleaned with bleach or ethanol and deionized water prior to use and after handling each specimen. Pipettes Pipettes are cleaned on a regular basis (see Chapter 2). Pipette tips Centrifuges Microcentrifuge tubes Only barrier (plug) tips are used when pipetting DNA samples. Tips are changed between each sample. The centrifuges used in the DNA analysis section are cleaned routinely with ethanol. If a spill occurs, the surfaces are wiped with ethanol followed by 1% bleach solution prior to further use. All microcentrifuge tubes are opened with a decapping tool only. Sterile tubes are used exclusively. Page 1 of 5 All printed copies are uncontrolled. This document printed on 7/22/215

2 , Continued Thermal cyclers Extraction fume hood Biosafety cabinet DNA spills Extraction and set-up area benchtops Amplification/ typing room benchtops Disposable mops The thermal cyclers are wiped down routinely with ethanol. Areas to be cleaned are the heating block and outer surfaces of the instrument. Individual wells are cleaned as needed. As with the bench top work area, the hood used for phenol/chloroform extractions is periodically cleaned with 1% bleach or ethanol. Clean absorbent disposable pads are placed over the cleaned area prior to use. After each use, the biosafety cabinet is cleaned with water and then with ethanol. Pipettes and decappers used in the biosafety cabinet are wiped down with ethanol. A DNA spill, either extracted or amplified, requires immediate attention. The spill is wiped up with absorbent paper wipes. The affected area is then cleaned with a 1% bleach solution by flooding the area with bleach and allowing it to stand for at least 1 minutes before wiping the liquid up with absorbent paper. All working bench top surfaces are thoroughly cleaned with ethanol or bleach before beginning a new case. Clean absorbent disposable pads are placed on the bench top work area prior to use. All working benchtop surfaces are periodically wiped down with water and then with ethanol. After wiping, a clean absorbent disposable pad should be placed on the bench top. This pad should be discarded into the waste containers in the amplification/typing room after each typing set. When work is concluded in these rooms, all equipment used should be wiped down with ethanol (for example, pipettes and centrifuges). Disposable mops are used to mop the floor in the amplification/typing room. The disposable mops are discarded and removed from the laboratory in a plastic garbage bag. They are disposed of in the outside trash bin. Page 2 of 5 All printed copies are uncontrolled. This document printed on 7/22/215

3 , Continued Glassware cleaning The general procedure for cleaning glassware is as follows. Step Action 1 Wash glassware with laboratory detergent. Heavily soiled items may be soaked for several hours. 2 Rinse with tap water. 3 Re-rinse with deionized water. 4 Allow to dry. NOTE: For many applications, washing with a mild detergent will remove grease and oil. When more rigorous cleaning is needed, organic solvents (for example, alcohol or acetone) may be used. This should then be followed by the regular cleaning procedures described above. The operating temperature of the autoclave is checked yearly by the manufacturer. The operating temperature of the autoclave is checked semi-annually by the laboratory staff. Results are recorded on the Temperature Verification form in the Equipment Calibration and Maintenance logbook located in the extraction area. The desired temperature range should be within 5 C of the displayed temperature. If the temperature is out of the desired range, refer to the instruction manual. For instructions for use, refer to the autoclave operating manual located in the buffer preparation area. Page 3 of 5 All printed copies are uncontrolled. This document printed on 7/22/215

4 , Continued glassware and plastics solutions Inert waste disposal Glass containers may be autoclaved. Only autoclave appropriate types of plastics (for example, polypropylene, polymethylpentene, polypropylene copolymer). Polycarbonate may be autoclaved but the cycle should be limited to 2 minutes at 121 C. All items should be carefully cleaned before autoclaving. Set the cap or closure on top of the container without engaging the threads. Attach an autoclave indicator to the container. Small items may be autoclaved inside a beaker covered with foil. Do not autoclave detergents or wetting solutions (for example, SDS). Do not autoclave solutions containing DTT. All containers should be carefully cleaned before autoclaving. Do not fill containers with solutions to more than 75% of capacity. Set the cap or closure on top of the container without engaging the threads. Attach an autoclave indicator to the container. DNA extraction and PCR set-up areas: Inert waste is emptied weekly into a main trash receptacle in the biology laboratory. Amplification/typing room: Inert waste is emptied weekly. The trash is placed in the hallway outside of the Biology Laboratory and picked up by the janitor to be disposed of in containers located in the large trash bin outside. Page 4 of 5 All printed copies are uncontrolled. This document printed on 7/22/215

5 , Continued Biohazard waste disposal DNA Extraction and PCR set-up areas: Biohazard waste is removed and transported to the biohazard waste containers located in the loading dock area for pick-up by the waste hauler weekly. Page 5 of 5 All printed copies are uncontrolled. This document printed on 7/22/215