Hello,
We tried the Illumina Truseq DNA preparation and followed by Illumina or Nimbelgen exon capture procedure. But we are having problems with out library preparation.
The input is 1 ug genomic DNA (quantified by Qubit 2.0). We exactly followed the illumina protocol, but the libraries we got after enrichment PCR is about 250-500ng total DNA (Bioanalyzer). This is not enough for later exon capture by either Illumina or Nimbelgen procedure.
Illumina told that they do not guarantee to get more than 1 ug library after the PCR, and as long as total amount is more than 500ng, Illumina exon capture should be fine. If I want to get 1 ug DNA, which is required by Nimbelgen protocol, I should do 2 PCR and pool them together.
Is it possible anyone here can help us out?
Thank you very much!
We tried the Illumina Truseq DNA preparation and followed by Illumina or Nimbelgen exon capture procedure. But we are having problems with out library preparation.
The input is 1 ug genomic DNA (quantified by Qubit 2.0). We exactly followed the illumina protocol, but the libraries we got after enrichment PCR is about 250-500ng total DNA (Bioanalyzer). This is not enough for later exon capture by either Illumina or Nimbelgen procedure.
Illumina told that they do not guarantee to get more than 1 ug library after the PCR, and as long as total amount is more than 500ng, Illumina exon capture should be fine. If I want to get 1 ug DNA, which is required by Nimbelgen protocol, I should do 2 PCR and pool them together.
Is it possible anyone here can help us out?
Thank you very much!
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