Hello
i just want to know if it is possible to check the successful AFLP selective amplification step using normal agarose gel 1%.
i tried it on 1% gel at 94 v for 20 mints but i didn't got any smear or band, i am using ECOR1 and MSE1 primer combinations but i only use one combination and non fluorescent ones to check for the first time.
Annealing temp is 65 and reducing each step 1oc till 56 (touch down PCR) but my primers has low anneling temp 45 and 48.
Hi, you can verify the success of AFLP selective amplificaiton using agarose electrophoresis. The more selective nucleotids of primers, the less bands less than 1000 bp or smear will be seen in the agarose gel.
Hi Nisar,
I agree with Zhanju and Moradi.
I check the success of AFLP selective amplification on 2.5 agarose gel.
Yes the AFLP selective amplification can be seen on agarose gel and as Zhanji Liu said a smear will appear. The appearnce of smear within range of 400-700 with higher intensity towards 500bp reveals that you are going to get a quality data.
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