[QUOTEThe only difference is the entry DNA, since they are amplicons, the concentration is a lot lower. On average around 200 ng, which is less than the recommended 1µg-5µg input. Should I alter the quantities of the reagents? ][/QUOTE]
I am not aware of any current kit requiring 1-5 ug input DNA, although few years ago it was common for shotgun library prep kits. In Illumina's TruSeq Nano DNA kit which requires 100-200 ng input depending on intended insert size, after size selection, end repair and A tailing around 20-50 ng DNA is usually remains for ligation step. With that input amount one can prepare good libraries with half reactions. You can use 20-30 ng of amplicons with half reactions from A tailing step onward. I would recommend cleaning up your amplicons before A tailing for good results.
I am not aware of any current kit requiring 1-5 ug input DNA, although few years ago it was common for shotgun library prep kits. In Illumina's TruSeq Nano DNA kit which requires 100-200 ng input depending on intended insert size, after size selection, end repair and A tailing around 20-50 ng DNA is usually remains for ligation step. With that input amount one can prepare good libraries with half reactions. You can use 20-30 ng of amplicons with half reactions from A tailing step onward. I would recommend cleaning up your amplicons before A tailing for good results.
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