Hello,
Each time, I used QIAquick gel extraction kit to extract the size selection product, the OD 260/230 is extremely weird (very low, close to 0), and 260/280 is a little bit higher (2.0-2.1). I found the absorption peak was moved from 260nm to 230nm, but the DNA should be extracted. I guess the remaining salt (such as EDTA) causes this, but I've tried to optimize the gel extraction step for several times (such as adding 6X volumes of QG, extending incubating time for QG, standing 5min after adding PE, ...), but they didn't work.
The colleagues at lab also have this problem, but it didn't affect their TOPO cloning or Sanger sequencing.
I have no idea if such weird ratio will affect some steps in the Illumina paired-end library preparation, such as Phusion PCR reaction or Cluster generation?
Anyone has such experience?
Many thanks,
SK
Each time, I used QIAquick gel extraction kit to extract the size selection product, the OD 260/230 is extremely weird (very low, close to 0), and 260/280 is a little bit higher (2.0-2.1). I found the absorption peak was moved from 260nm to 230nm, but the DNA should be extracted. I guess the remaining salt (such as EDTA) causes this, but I've tried to optimize the gel extraction step for several times (such as adding 6X volumes of QG, extending incubating time for QG, standing 5min after adding PE, ...), but they didn't work.
The colleagues at lab also have this problem, but it didn't affect their TOPO cloning or Sanger sequencing.
I have no idea if such weird ratio will affect some steps in the Illumina paired-end library preparation, such as Phusion PCR reaction or Cluster generation?
Anyone has such experience?
Many thanks,
SK
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