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Old 10-10-2012, 07:43 AM   #1
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Location: chicago

Join Date: Jan 2011
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Default TruSeq DNA 500bp library prep

Hello, we are planning to make a TruSeq library with an starting insert of 500bp and worried that this higher molarity might affect the subsequent TruSeq kit rxns. The max range recommended in the manuel is 300bp insert.

Would multiplying the reagents and volumes by (500/300 ) = 1.667 be the best course? Or start with .67ug of the 500bp gDNA?
Does anyone have experience making TruSeq libraries this size?

thanks, ap
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Old 10-10-2012, 08:28 AM   #2
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Location: Bethesda MD

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Er, your math is backwards. As the size increases, the molarity (number of molecules) per mass unit decreases.

Having said that, I doubt that a difference of less than 2X will have a profound effect on the library construction. But I recommend that you search the forum for the impact of insertion size on clustering and data quality.

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library generation, size factors, truseq dna prep kit

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