Bacterial Whole Genome Sequencing Library Preparation

October 13, 2017

University of Birmingham
School of Biosciences
Postdoctoral Research Fellow

Overall

Quality of Results

Ease-of-Optimization

What do these ratings mean?
Write a Review

Company:

New England Biolabs

Product Name:

NEBNext® Ultra™ II DNA Library Prep Kit for Illumina®

Catalog Number:

E7645S

I used NEBNext Ultra II DNA library prep kit for S. aureus genome sequencing using Illumina Miseq platform. Used 100 ng sheared DNA as input and got 4nM library yield with 10 PCR cycles. After sequencing, got 3.5 M reads with coverage of 97 %.

Experimental Design and Results Summary

Application

Bacterial whole genome sequencing

Starting Material

100 ng of sheared S. aureus genomic DNA

Protocol Overview

100 ng of sheared S. aureus genomic DNA in 50 microliter volume was used with NEBNext Ultra II DNA library prep kit. Followed standard and easy to follow protocol guidelines given in the kit manual. After ligation of adapters, performed 10 cycles of PCRs to amplify library suitable of Illumina Miseq platform. Quantified libraries using NEB library quantification kit and got 4 nM yield.

Tips

Always store kit contents in -20 degree freezer. Avoid frequent freeze thaw cycles.

Results Summary

Fast and easy to follow protocol of preparing DNA library for sequencing in Illumina platform. 3.5 million reads were obtained using this library prep kit with more than 90 percent reads mapping to genome.

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

Image Gallery

Summary

The Good

Simple to follow and easy protocol. Good thing is that it can used for low input samples such 500 picogram of sheared DNA.

The Bad

Nothing I came so far.

The Bottom Line

I regularly use this kit for bacterial whole genome sequencing since the kit really does the job and protocol and steps are simple and easy to follow.

Join the discussion