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Old 02-19-2016, 11:30 AM   #1
krabidou
Junior Member
 
Location: boston

Join Date: Feb 2016
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Default DNA library shoulder peak on bioanalyzer

Hi Everyone,

We recently just processed a few DNA libraries using the Illumina Truseq PCR free kit. When we ran them on Bioanalyzer, one of the libraries came out perfect, with a single peak centered around 500 bp. The others, however, have a secondary shoulder peak ( ~ 375 bp) attached to the main library curve. We have never had this happen to our previous libraries. Has anyone else seen this/know the underlying cause and how to avoid it? Thanks!
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Old 02-19-2016, 03:20 PM   #2
nucacidhunter
Jafar Jabbari
 
Location: Melbourne

Join Date: Jan 2013
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Default

Few points:

1- Chip is overloaded
2- If shoulder is not consistent among the samples it could be due to vibration during Chip run.
3- Is the perfect library concentration is lower than the others. If true it would indicate artefact of overloading Chip.
4- The trace is not similar to TruSeq PCR-free library profiles (350 or 550 bp insert)
5- If shoulder is real, could be the result of size selection step
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bioanalyzer, dna library prep, illumina, truseq dna

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