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Question: Stranded-specific library by dUTP method
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Hi all,

Regarding stranded-specific library preparation by dUTP method, as far as I know, the dUTP is used during the synthesis of the second strand, which will be degraded before PCR amplification step, so just the first strand is sequenced. What about the sequencing of the second strand (coding or sense strand)? I searched on the net a lot, but I didn't find a clear answer for it. Could you please kindly tell me about the second strand sequencing? it will be great if there is any schematic image.

Thank you

ADD COMMENTlink 3.3 years ago seta ♦ 1.2k • updated 3.3 years ago genomax 68k
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ADD COMMENTlink 3.3 years ago genomax 68k
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Thank you genomax2. However, the question still remains for me since it sounds that just the first strand of cDNA are sequenced and produced read 1. What about the sequencing of the second strand, which degraded by uracil DNA glycosylase before PCR amplification? Please clear me on this issue.

ADD REPLYlink 3.3 years ago
seta
♦ 1.2k
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DNA strands are complementary correct? Once you sequence one you get the other.

ADD REPLYlink 3.3 years ago
genomax
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oh, yes. Thank you and sorry

ADD REPLYlink 3.3 years ago
seta
♦ 1.2k

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