How much template is required for A-tailing of an amplified product?

23 September 2021 1 4K Report

I have amplified ~800kb fragment and want to proceed with A-tailing of this product. How much template I need for this size. It did not work when I used 100ng of template which was a gel excised PCR product.

Miguel Fernández-Niño

It seems that you have to use more. The recommended amount is 10–100 ng PCR product for each 100 bp length of the PCR product. Remember to purify your PCR first fron gel to remove the proof reading poly. :)

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Science method

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