60110 
NOTICES 
II-B-1. PI Level. 
II-B-l-a. Laboratory Practices. 
II-B-l-a-(l). Laboratory doors shall 
be kept closed while experiments are 
in progress. 
II-B-l-a-(2). Work surfaces shall be 
decontaminated daily, and immediate- 
ly following spills of organisms con- 
taining recombinant DNA molecules. 
II-B-l-a-(3). All biological wastes 
shall be decontaminated before dispos- 
al. Other contaminated materials, 
such as glassware, animal cages, and 
laboratory equipment, shall be decon- 
taminated before washing, reuse, or 
disposal. 
II-B-l-a-(4). Mechanical pipetting 
devices shall be used; pipetting by 
mouth is prohibited. 
II-B-l-a-(5). Eating, drinking, smok- 
ing, and storage of foods are not per- 
mitted in the laboratory area in which 
recombinant DNA materials are han- 
dled. 
II-B-l-a-(6). Persons shall wash 
their hands after handling organisms 
containing recombinant DNA mole- 
cules and when they leave the labora- 
tory. 
II-B-l-a-(7). Care shall be taken in 
the conduct of all procedures to mini- 
mize the creation of aerosols. 
II-B-l-a-(8). Contaminated materi- 
als that are to be decontaminated at a 
site away from the laboratory shall be 
placed in a durable leak-proof contain- 
er, which is closed before removal 
from the laboratory. 
II-B-l-a-(9). An insect an rodent 
control program shall be instituted. 
II-B-l-a-(lO). The use of laboratory 
gowns, coats, or uniforms is discretion- 
ary with the laboratory supervisor. 
II-B-l-a-(ll). Use of the hypoder- 
mic needle and syringe shall be avoid- 
ed when alternative methods are avail- 
able. 
II-B-l-a-(12). The laboratory shall 
be kept neat and clean. 
II-B-l-b. Containment Equipment. 
Special containment equipment is not 
required at the PI level. 
II-B-l-c. Special Laboratory Design. 
Special laboratory design is not re- 
quired at the pi level. 
II-B-2. P2 Level. 
II-B-2-a. Laboratory Practices. 
II-B-2-a-( 1 ). Laboratory doors shall 
be kept closed while experiments are 
in progress. 
II-B-2-a-(2). Work surfaces shall be 
decontaminated daily, and immediate- 
ly following spills of organisms con- 
taining recombitant DNA molecules. 
II-B-2-a-(3). All laboratory wastes 
shall be steam-sterilized (autoclaved) 
before disposal. Other contaminated 
materials such as glassware, animal 
cages, laboratory equipment, and ra- 
dioactive wastes shall be decontami- 
nated by a means demonstrated to be 
effective before washing, reuse, or dis- 
posal. 
FEDERAL 
II-B-2-a-(4). Mechanical pipetting 
devices shall be used; pipetting by 
mouth is prohibited. 
II-B-2-a-(5). Eating, drinking, smok- 
ing, and storage of food are not per- 
mitted in the laboratory area in which - 
recombinant DNA materials are han- 
dled. 
II-B-2-a-(6). Persons shall wash 
their hands after handling organisms 
containing recombinant DNA mole- 
cules and when they leave the labora- 
tory. 
II-B-2-a-(7). 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 in- 
oculation loop or needle so that it 
splatters, and forceful ejection of 
fluids from pipettes or syringes shall 
be avoided. 
II-B-2-a-(8). Contaminated materi- 
als that are to be steam sterilized (au- 
toclaved) or decontaminated at a site 
away from the laboratory shall be 
placed in a durable leak-proof contain- 
er, which is closed before removal 
from the laboratory. 
II-B-2-a-(9). Only persons who have 
been advised of the nature of the re- 
search being conducted shall enter the 
laboratory. 
II-B-2-a-(10). The universal bioha- 
zard sign shall be posted on all labora- 
tory access doors when experiments 
requiring P2 containment are in prog- 
ress. Freezers and refrigerators or 
other units used to store organisms 
containing recombinant DNA mole- 
cules shall also be posted with the uni- 
versal biohazard sign. 
II-B-2-a-(ll). An insect and rodent 
control program shall be instituted. 
II-B-2-a-(12). 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 lo- 
cated. 
II-B-2-a-(13). Animals not related to 
the experiment shall not be permitted 
in the laboratory. 
II-B-2-a-(14). Use of the hypoder- 
mic needle and syringe shall be avoid- 
ed when alternative methods are avail- 
able. 
II-B-2-a-(15). The laboratory shall 
be kept neat and clean. 
II-B-2-a-(16). Experiments of lesser 
biohazard potential can be carried out 
concurrently in carefully demarcated 
areas of the same laboratory. 
II-B-2-b. Containment Equipment 
Biological safety cabinets[20] shall be 
used to contain aerosol-producing 
equipment, such as blenders, lyophi- 
lizers, sonicators, and centrifuges, 
when used to process organisms con- 
taining recombinant DNA molecules, 
except where equipment design pro- 
vides for containment of the potential 
aerosol. For example, a centrifuge may 
REGISTER, VOL. 43, NO. 247— FRIDAY, DECEMBER 
[ 32 ] 
be operated in the open if a sealed 
head or safety centrifuge cups are 
used. 
II-B-2-c. Special Laboratory Design. 
An autoclave for sterilization of wastes 
and contaminated materials shall be 
available in the same building in 
which organisms containing recombin- 
ant DNA molecules are used. 
II-B-3. P3 Level. 
II-B-3-a. Laboratory Practices. 
II-B-3-a-(l). Laboratory doors shall 
be kept closed while experiments are 
in progress. 
II-B-3-a-(2). Work surfaces shall be 
decontaminated following the comple- 
tion of the experimental activity, and 
immediately following spills of organ- 
isms confining recombinant DNA mol- 
ecules. 
II-B-3-a-(3). All laboratory wastes 
shall be steam-sterilized (autoclaved) 
before disposal. Other contaminated 
materials, such as glassware, animal 
cages, laboratory equipment, and ra- 
dioactive wastes, shall be decontami- 
nated by a method demonstrated to be 
effective before washing, reuse, or dis- 
posal. 
II-B-3-a-(4). Mechanical pipetting 
devices shall be used; pipetting by 
mouth is prohibited. 
II-B-3-a-(5). Eating, drinking, smok- 
ing, and storage of food are not per- 
mitted in the laboratory area in which 
recombinant DNA materials are han- 
dled. 
II-B-3-a-(6). Persons shall wash 
their hands after handling organisms 
containing recombinant DNA mole- 
cules and when they leave the labora- 
tory. 
II-B-3-a-(7). 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 in- 
oculation loop or needle , so that it 
splatters, and forceful ejection of 
fluids from pipettes or syringes shall 
be avoided. 
II-B-3-a-(8). Contaminated materi- 
als that are to be steam-sterilized (au- 
toclaved) or decontaminated at a site 
away from the laboratory shall be 
placed in a durable leak-proof contain- 
er, which is closed before removal 
from the laboratory. 
II-B-3-a-(9). Entry into the labora- 
tory shall be . through a controlled 
access area. Only persons who have 
been advised of the nature of the re- 
search 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. 
22, 1978 
