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Old 07-28-2015, 09:02 AM   #2
donquijotes
Junior Member
 
Location: Michigan

Join Date: Jul 2015
Posts: 7
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Hi sudders,

From my very limited experience with random barcodes, I've heard that this bias could be due to 2 reasons.
a) Synthesis bias. Some oligo companies suggest manually mixing the 4 bases when they do random synthesis. One rep once told me that they have seen 20-30% variation/bias in base incorporation with automatic mixing.
b) Ligation bias. Some bases/sequences ligate less efficiently to your library.

BTW, I would like to design a 5 random barcode and have it in both ends of my DNA library. Do you know of any open source pipeline out there that will help me get rid of the PCR duplicates using both paired end barcodes? (1 million combinations total)
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