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Hello to everyone. I am new in this forum.
I am working on a library for reduced representation bisulfite sequencing (RRBS). The protocol I have mentioned the adaptor sequences which comprised with methylated C, an overhanded T that can ligate with the target sequence A overhanded. The last T is also linked with phosphorothiate group which can protect them from 3->5 exonuclease activity.
I noticed that the two sequences of the adaptor can only anneal for half of these sequences, which, I believe, make it sufficient to ligate with the target sequence. But I am still wondering why don't we make two adaptor sequences with full complement to each other?
I am working on a library for reduced representation bisulfite sequencing (RRBS). The protocol I have mentioned the adaptor sequences which comprised with methylated C, an overhanded T that can ligate with the target sequence A overhanded. The last T is also linked with phosphorothiate group which can protect them from 3->5 exonuclease activity.
I noticed that the two sequences of the adaptor can only anneal for half of these sequences, which, I believe, make it sufficient to ligate with the target sequence. But I am still wondering why don't we make two adaptor sequences with full complement to each other?