Quantification of Bacterial DNA Sequencing Libraries for Illumina Miseq Platform

University of Birmingham
School of Biosciences
Postdoctoral Research Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

KAPA Biosystems

Product Name:

KAPA Library Quantification Complete Kit for Illumina

Catalog Number:

KK4824

I have used KAPA Library quantification Kit for Illumina to quantify my DNA sequencing libraries. The kit is based on SYBR green based detection of DNA libraries and performs well with my samples in standard protocols as mentioned in the instruction manual. I have used this qPCR master mix in Agilent's Mx3500P suit and got consistent results on library quantification which eventually gave my optimal cluster amplifications in Miseq.

Experimental Design and Results Summary

Application

Quantification of DNA libraries for Illumina Miseq

Starting Material

Bacterial DNA libraries prepared with NEBNext Ultra DNA library prep kit.

Protocol Overview

Protocol is pretty simple and followed the instructions that come with the kit manual. First, prepare DNA standards which come with the kit and place them in 96 well microplates (each in 20 microliter reaction). Then prepare series of 10 fold dilutions on your DNA libraries and place them in the same 96 well microplate in 20 microliter reaction according to kit's instructions. qPCR cycling conditions and data analysis were performed as per instructions of the kit manual.

Tips

Do not use the kit if it is expired (as it says on the kit label)

Results Summary

Got consistent linear standard curve generation for DNA standards. My libraries were in the range of 50 nM concentration after qPCR quantification using KAPA quant kit.

Additional Notes

None

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Summary

The Good

Kit protocol is simple to perform. No complications.

The Bad

None

The Bottom Line

Highly recommended for quantification of DNA libraries intended for use in Illumina sequencing platform.

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