I have some question about Colony PCR:
1. pGLO causes the transformed colonies to glow. Why do we use ampicillin selection plates when we can identify transformed bacteria optically.
A. ampicillin increases GFP expression
B. ampicillin reduces competition from untransformed bacteria
C. ampicillin is not necessary
D. ampicillin is necessary because we are not using aseptic/sterile technique
2. When picking colonies for 'Colony PCR',
A. take a large streak from the densest part of the bacterial lawn
B. choose the center of a small colony
C. dig the agarose from underneath a small colony
D. combine 2-3 small colonies
3. In colony PCR a good positive control would be ....
A. the plasmid from the transformation
B. a section of the bacterial lawn
C. a liquid culture of the same colony
D. a master mix with no PCR primer
4. If colony PCR shows a band of the correct size, when is sequencing required?
A. to make sure the correct gene is being expressed
B. to determine orientation of the gene
C. to confirm point mutations
D. both B and C
5. How do you grow the bacteria from the PCR positive colony after colony PCR?
A. You can't, PCR kills the bacteria. Just pick another colony
B. Streak the PCR products on an AMP agarose plate, the bacteria will grow just fine.
C. Don't add the entire colony to the PCR master mix
D. Re-transform bacteria assuming the same transformation efficiency
I have some question about Colony PCR:
1. pGLO causes the transformed colonies to glow. Why do we use ampicillin selection plates when we can identify transformed bacteria optically.
A. ampicillin increases GFP expression
B. ampicillin reduces competition from untransformed bacteria
C. ampicillin is not necessary
D. ampicillin is necessary because we are not using aseptic/sterile technique
2. When picking colonies for 'Colony PCR',
A. take a large streak from the densest part of the bacterial lawn
B. choose the center of a small colony
C. dig the agarose from underneath a small colony
D. combine 2-3 small colonies
3. In colony PCR a good positive control would be ....
A. the plasmid from the transformation
B. a section of the bacterial lawn
C. a liquid culture of the same colony
D. a master mix with no PCR primer
4. If colony PCR shows a band of the correct size, when is sequencing required?
A. to make sure the correct gene is being expressed
B. to determine orientation of the gene
C. to confirm point mutations
D. both B and C
5. How do you grow the bacteria from the PCR positive colony after colony PCR?
A. You can't, PCR kills the bacteria. Just pick another colony
B. Streak the PCR products on an AMP agarose plate, the bacteria will grow just fine.
C. Don't add the entire colony to the PCR master mix
D. Re-transform bacteria assuming the same transformation efficiency
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