Ran AFB PCR using the same DNA template previously used in the attempted 16S amplification attempt. The results were less than desirable.
The top lanes are the AFB amplicons and the bottom is the 16S amplicons. Lane labels are below the images. The two gels are the same, the one on the right it just overexposed so I could see it more clearly here.
| Lane 1 | Lane 2 | Lane 3 | Lane 4 | Lane 5 | Lane 6 | Lane 7 | Lane 8 |
| Hi Lo | 2a | 22a | 8a | 63a | Q | RT | Pl |
A volume of 5 uL of amplicon and 2 uL of EZ vision were loaded into each of the lanes. Not all the isolates amplified when using the AFB primers. All of them them except the roach trachea "RT" isolate should have! The positive control, P. larvae 9545 "Pl", checked out though. As you can also see, there was no bands that appeared using the 16S primers, however there was a lot of DNA in the wells. Perhaps I am overloading the lanes?
Working on a solution to these issues...
//EWW
//EWW
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