Hi guys,
When prerparing an illumina RNA-seq library, i found this instruction:
In secondary strand synthesis the DNA polymerase use the first-stranded cDNA as template and the RNase H digested mRNA as primers to synthesize DNA. Some where the 5' RNA could be replaced and the fragments of double stranded cDNA could be ligated by the E.coli DNA ligase in the reaction.
I believe there's no DNA ligase added in this reaction for my library, meanwhile i dont think DNA poly has ligase activity. So i'll appreciate anyone who helps me understand better about how this works.
Thanks.
When prerparing an illumina RNA-seq library, i found this instruction:
In secondary strand synthesis the DNA polymerase use the first-stranded cDNA as template and the RNase H digested mRNA as primers to synthesize DNA. Some where the 5' RNA could be replaced and the fragments of double stranded cDNA could be ligated by the E.coli DNA ligase in the reaction.
I believe there's no DNA ligase added in this reaction for my library, meanwhile i dont think DNA poly has ligase activity. So i'll appreciate anyone who helps me understand better about how this works.
Thanks.
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