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Old 02-07-2014, 11:58 AM   #1
Daytwa
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Location: Detroit

Join Date: Dec 2010
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Default Innaccurate Kapa Quant Standards

On my campus two labs, both with very extensive experience in NGS, have reported that recently purchased Kapa qPCR standards have under-estimated library concentrations. Both sets of libraries were quantified following two similar but lab dependent protocols (one uses the full kit the other uses just the standard and an off the shelf taq-mix) and then diluted to 2 nM and the dilutions were then quantified. Typical amounts of library (10 and 12 pmol/lane; the latter exceeds Illumina's rec. but the group swears by it) were run on HiSeq 8-lane flowcells but produced ~50% of the expected reads (~4x10^8/lane). Typically these labs see 6 - 8 x10^8 reads/lane. One lane contained a sample which was being re-run and was quantified months before with a different lot of Kapa. This recent run of this sample produced the expected number of reads. There was no issue with the quality scores any samples and the data are fine (just lower coverage). The library quantifications were double checked using the suspect Kapa and it doesn't appear to be anything to do with the PCRs.

The dates of purchase and shipping/receiving of the suspect Kapa kits are very similar (Nov 2013) but the lot numbers are different. Has anyone had any trouble with Kapa standards recently?

UPDATE: We spoke to Kapa and they are having problems with lots. I would call ahead before running anything.

Last edited by Daytwa; 02-07-2014 at 12:07 PM. Reason: New News
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Old 02-11-2014, 01:09 PM   #2
AdrianaGeldart
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Location: Boston, MA

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Default

I am a Kapa Biosystems Application Scientist and just wanted to clarify the above post. When a customer calls into Kapa Biosystems Support with a complaint regarding a product's performance, we ask for supporting data and the product lot number(s). This is standard procedure for all complaint inquiries, and does not imply that we have an issue with any lots. I would like to reassure everyone that there is no systemic issues with our KAPA Library Quantification DNA Standards. If a customer is concerned about their product, please feel free to contact Kapa Biosystems directly at support@kapabiosystems.com or 855-527-2246. Thank you.
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