E. coli Genome Sequencing Using Nextera XT DNA Library Prep Kit

University of Birmingham
School of Biosciences
Postdoctoral Research Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

Illumina

Product Name:

Nextera XT DNA Library Preparation Kit

Catalog Number:

FC-131-1024

We routinely do bacterial isolates genome sequencing and for that we use Nextera XT DNA library prep kit. Usually we get very good quality library at 50 to 100 nM concentration which are then equally pooled with indexed libraries and finally 20 pM was loaded onto Miseq. Over 90 percent of reads had Q score more than 30.

Experimental Design and Results Summary

Application

Bacterial genome sequencing

Starting Material

E. coli genomic DNA

Protocol Overview

Bacterial isolates were used for genomic DNA preparation. Then we use Nextera Index Kit to introduce indexing (multiplexing the isolates). Library preparation is very simple and just follow standard protocol guide form the kit. Finally we load 20 pM denatured libraries onto Miseq platform for sequencing.

Tips

None

Results Summary

All the libraries prepared were of good quality as checked by Agilent bioanalyser using DNA chip. Over 90 percent of reads pass the filter and had a Q score greater than 30.

Additional Notes

None

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Summary

The Good

Very easy to perform protocol. Entire protocol can be finished in less than 3 hrs with hands-on-time less than 30 minutes.

The Bad

None

The Bottom Line

Highly recommended for large scale genome sequencing different bacterial isolates.

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