24570 Federal Register 
container, which is closed before 
removal from the laboratory. 
Appendix G-II-A-l-i. An insect and 
rodent control program shall be 
instituted. 
Appendix G — II— A— 1 — j. The use of 
laboratory gowns, coats, or uniforms is 
discretionary with the laboratory 
supervisor. 
Appendix G-Il-A-l-k. Use of the 
hypodermic needle and syringe shall be 
avoided when alternative methods are 
available. 
Appendix G-II-A-1-1. The laboratory 
shall be kept neat and clean. 
Appendix G-II-A-2. Containment 
Equipment. Special containment 
equipment is not required at the Pi level. 
Appendix G-II-A-3. Special 
Laboratory Design. Special laboratory 
design is not required at the Pi level. 
Appendix G-1I-B. P2 Level. 
Appendix G-II-B-1. Laboratory 
Practices. 
Appendix G-II-B-l-a. Laboratory 
doors shall be kept closed while 
experiments are in progress. 
Appendix G-II-B-l-b. Work surfaces 
shall be decontaminated daily, and 
immediately following spills of 
organisms containing recombinant DNA 
molecules. 
Appendix G-II-B-l-c. All laboratory 
wastes shall be steam-sterilized 
[autoclaved) before disposal. Other 
contaminanted materials such as 
glassware, animal cages, laboratory 
equipment, and radioactive wastes shall 
be decontaminated by a means 
demonstrated to be effective before 
washing, reuse, or disposal. 
Appendix G-Il-B-l-d. Mechanical 
pipetting devices shall be used; pipetting 
by mouth is prohibited. 
Appendix G-Il-B-l-e. Eating, 
drinking, smoking, and storage of food 
are not permitted in the laboratory area 
in which recombinant DNA materials 
are handled. 
Appendix G — II— 13 — 1— f. Persons shall 
wash their hands after handling 
organisms containing recombinant DNA 
molecules and when they leave the 
laboratory. 
Appendix G— Il-B-l-g. Care shall be 
exercised to minimize the creation of 
aerosols. For example, manipulations 
such as inserting a hot inoculating loop 
or needle into a culture, flaming an 
inoculation loop or needle so that it 
splaters, and forceful ejection of fluids 
from pipettes or syringes shall be 
avoided. 
Appendix G-ll-B-l-h. Contaminated 
materials that are to be steam sterilized 
(autoclaved) or decontaminated at a site 
away from the laboratory shall be 
placed in a durable leak-proof container, 
/ Vol. 48, No. 10G / Wednesday, June 1, 1983 / Notices 
which is closed before removal from the 
laboratory. 
Appendix G — II— E3— 1— i. Only persons 
who have been advised of the nature of 
the research being conducted shall enter 
the laboratory. 
Appendix G-II-B-l-j. The universal 
biohazard sign shall be posted on all 
laboratory access doors when 
experiments requiring P2 containment 
are in progress. Freezers and 
refrigerators or other units used to store 
organisms containing recombinant DNA 
molecules shall also be posted with the 
universal biohazard sign. 
Appendix G— II-B-l-k. An insect and 
rodent control program shall be 
instituted. 
Appendix G— II— B— 1— 1. The use of 
laboratory gowns, coats, or uniforms is 
required. Laboratory clothing shall not 
be worn to the lunch room or outside of 
the building in which the laboratory is 
located. 
Appendix G-II-B-l-m. Animals not 
related to the experiment shall not be 
permitted in the laboratory. 
Appendix G-II-B-l-n. Use of the 
hypodermic needle and syringe shall be 
avoided when alternative methods are 
available. 
Appendix G-II-B-l-o. The laboratory 
shall be kept neat and clean. 
Appendix G-II-B-l-p. Experiments of 
lesser biohazard potential can be 
carried out concurrently in carefully 
demarcated areas of the same 
laboratory. 
Appendix G-II-B-1-2. Containment 
Equipment. Biological safety cabinets 
[12] shall be used to contain aerosol- 
producing equipment, such as blenders, 
lyophilizers, sonicators, and centrifuges, 
when used to process organisms 
containing recombinant DNA molecules, 
except where equipment design 
provides for containment of the 
potential aerosol. For example, a 
centrifuge may be operated in the open 
if a sealed head or safety centrifuge 
cups are used. 
Appendix G-II-B-1-3. Special 
Laboratory Design. An autoclave for 
sterilization of wastes and contaminated 
materials shall be available in the same 
building in which organisms containing 
recombinant DNA molecules are used. 
Appendix G-II-C. P3 Level. 
Appendix G-Il-C-1. Laboratory 
Practices. 
Appendix G-II-C-l-a. Laboratory 
doors shall be kept closed while 
experiments are in progress. 
Appendix G-II-C-l-b. W'ork surfaces 
shall be decontaminated following the 
completion of the experimental activity, 
and immediately following spills of 
organisms containing recombinant DNA 
molecules. 
Appendix G-II-C-l-c. All laboratory 
wastes shall be steam-sterilized 
(autoclaved) before disposal. Other 
contaminated materials, such as 
glassware, animal cages, laboratory 
equipment, and radioactive wastes, 
shall be decontaminated by a method 
demonstrated to be effective before 
washing, reuse, or disposal. 
Appendix G-11-C-l-d. Mechanical 
pipetting devices shall be used; pipetting 
by mouth is prohibited. 
Appendix G-11-C-l-e. Eating, 
drinking, smoking, and storage of food 
are not permitted in the laboratory area 
in which recombinant DNA materials 
are handled. 
Appendix G-II-C-l-f. Persons shall 
wash their hands after handling 
organisms containing recombinant DNA 
molecules and when they leave the 
laboratory. 
Appendix G-II-C-l-g. Care shall be 
exercised to minimize the creation of 
aerosols. For example, manipulation 
such as inserting a hot inoculating loop 
or needle into a culture, flaming an 
inoculation loop or needle so that it 
splatters, and forceful ejection of fluids 
from pipettes or syringes shall be 
avoided. 
Appendix G-II-C-l-h. Contaminated 
materials that are to be steam-sterilized 
(autoclaved) or decontaminated at a site 
away from the laboratory shall be 
placed in a durable leak-proof container, 
which is closed before removal from the 
laboratory. 
Appendix G — II— C— 1— i. Entry into the 
laboratory shall be through a controlled 
access area. Only persons who have 
been advised of the nature of the 
research being conducted shall enter the 
controlled access area. Only persons 
required on the basis of program or 
support needs shall be authorized to 
enter the laboratory. Such persons shall 
be advised of the nature of the research 
being conducted before entry, and shall 
comply with all required entry and exit 
procedures. 
Appendix G— II— C— 1— j. Persons under 
16 years of age shall not enter the 
laboratory. 
Appendix G-Il-C-l-k. The universal 
biohazard sign shall be posted on the < 
controlled access area door and on all 
laboratory doors when experiments 
requiring P3-level containment are in 
progress. Freezers and refrigerators or 
other units used to store organisms 
containing recombinant DNA molecules 
shall also be posted with the universal J 
biohazard sign. 
rodent control program shall be 
instituted. 
[ 150 ] 
