Hi,
I am working with an Illumina Genome Analyser GaIIx. I am new at this, and I am studying to produce directional libraries for RNA-Seq. But I have some doubts about what I read in the following article referring to Illumina RNA ligation protocol:
Illumina decaps the RNA with TAP, fragments the RNA with Sodium citrate, then 3' dephosphorylates (with Antartic phosphatase) and then phosphorylates at its 5' end (with T4 polynucleotide kinase) . It don't understand why it does that, because as far as I understand, after decapping, RNA is monophosphorylated on its 5' end so it would be ready for adapter ligation...
Thank you very much for your help,
Pilar
I am working with an Illumina Genome Analyser GaIIx. I am new at this, and I am studying to produce directional libraries for RNA-Seq. But I have some doubts about what I read in the following article referring to Illumina RNA ligation protocol:
Illumina decaps the RNA with TAP, fragments the RNA with Sodium citrate, then 3' dephosphorylates (with Antartic phosphatase) and then phosphorylates at its 5' end (with T4 polynucleotide kinase) . It don't understand why it does that, because as far as I understand, after decapping, RNA is monophosphorylated on its 5' end so it would be ready for adapter ligation...
Thank you very much for your help,
Pilar
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