Repeated PCR of the honey isolates using the AFB primers. Ran 1% gel, 10 uL amplicon DNA + 2 uL EZ vision dye. Below is an image of the gel ran at 100 volts for 60 minutes.
| Lane 1 | Lane 2 | Lane 3 | Lane 4 | Lane 5 | Lane 6 | Lane 7 | Lane 8 |
| Hi Lo | 8a | 2a | 22a | 63a | Q | R. Trach | 9545 |
The template DNA used in this reaction is the same that was used in the previous PCR using the AFB primers only I used half as much as before. The positive control, P. larvae 9545, amplified using these P. larvae specific primers. The roach trachea (lane 7) served as a negative control as it has not amplified using these primers in the past and is highly suspected to a Bacillus spp. This time with the honey isolates you can clearly see the bands and there is minimal DNA left in the wells (no smearing). All of them amplifed except for 8a (Lane 2), which has amplified in the past indicating that it is indeed P. larvae . However, you can see that there is something in the well of this lane and it could be that I still used too much DNA for that isolate. In the future, I will only use half the amount of DNA that I have been using when setting up my PCR reactions.
//EWW
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