Hi all,
I'm preparing RAD sequencing library with a protocol inspired by Etter et al, except that I use Amplicon Beads instead of a gel.
Before sending to the sequencing platform I make a High sensitivity DNA chip (AGilent machine) to check the size of my fragment (normally between 300-800 base pairs).
On most of my samples I got a big peak at about 3000 base pairs, something that I don't understand.
Firts I thought it was the beads. But, this peak appear only after the PCR step, it is not there before PCR or either after sonication.
Is there anyone here that had the same problem ?
Also, some others people had the same problem than I with a different protocol, the GBS one (Elshire et al), but the same machine (Agilent).
Thanks for your help,
Laura
I'm preparing RAD sequencing library with a protocol inspired by Etter et al, except that I use Amplicon Beads instead of a gel.
Before sending to the sequencing platform I make a High sensitivity DNA chip (AGilent machine) to check the size of my fragment (normally between 300-800 base pairs).
On most of my samples I got a big peak at about 3000 base pairs, something that I don't understand.
Firts I thought it was the beads. But, this peak appear only after the PCR step, it is not there before PCR or either after sonication.
Is there anyone here that had the same problem ?
Also, some others people had the same problem than I with a different protocol, the GBS one (Elshire et al), but the same machine (Agilent).
Thanks for your help,
Laura
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