High Quality DNA Extraction for Long Read Sequencing

June 18, 2020

BlueskyReddit
University of Birmingham
School of Biosciences
Postdoctoral Research Fellow

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Company:

Revolugen

Product Name:

Revolugen Fire Monkey HMW DNA Extraction Kit

Catalog Number:

REV010

Getting good quality genomic DNA with high molecular weight with longer length is essential to get good quality sequencing reads from long read sequencing platform. Using Revolugen's Fire Monkey HMW DNA extraction kit, greater than 100 kb genomic DNA were extracted from bacterial samples,which indeed yielded high quality long read sequencing data from Nanopore sequencing. N50 of contigs from such DNA were 25-30 Kb length.

Experimental Design and Results Summary

Application

High quality DNA extraction for long read sequencing by Nanopore sequencing chemistry

Starting Material

1 mL of overnight grown bacterial cultures

Tips

Do not use more than 10^8 cells per column for DNA extraction

Results Summary

I used gram negative bacterial cultures for high quality with high molecular weight DNA extraction for long read sequencing by Nanopore sequencing platform. 1 mL overnight grown cultures were used for extraction according to kit's instruction protocol. Finally eluted genomic DNA in 180 microliter nuclease free water. This gave me 100 ng/microliter DNA with 260:280 ratio of 1.8. When extracted DNA was analysed in 0.6% agarose gel Electrophoresis, length of the extracted genomic DNA were greater than 100 kb in length, which is ideal for long read sequencing.

DOI or PMID #

N/A

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Summary

The Good

Simple user friendly protocol. Less hands on time.

The Bad

Nothing bad about this kit.

The Bottom Line

Highly recommended kit for high quality high molecular weight DNA for long read and other types of sequencing. Other available kits in the market are nowhere near to this Revolugen kit.

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