Quantification of Bacterial DNA

March 04, 2016

University of Birmingham
School of Biosciences
Postdoctoral Research Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

Thermo Fisher Scientific

Product Name:

Qubit dsDNA HS assay kit

Catalog Number:

Q32851

Bacterial DNA intended for NEB HiFi assembly and preparation of next generation sequencing was quantified by using Qubit dsDNA HS assay kit. Quantification was very specific to double stranded DNA in the samples since the fluorescent dye binds specifically to dsDNA. Using 1:200 dilution of neat DNA samples, I have gotten 50 ng/microliter of DNA samples.

Experimental Design and Results Summary

Application

Quantification of bacterial DNA for cloning and NGS

Starting Material

Bacterial DNA

Protocol Overview

Bacterial DNA was diluted into 1:5 using nuclease free water. Qubit dsDNA working buffer and standards were prepared as per manufacturer's instructions. Then diluted DNA samples were used at 1:200 dilution with working buffer. Standards were measured first then followed by samples in Qubit 2.0 Fluorimeter

Tips

If the samples reading fall out of the standard curve, dilute the neat DNA samples and then use diluted samples for Qubit fluorimetry.

Results Summary

Quantification results across three biological repeats ware consistent. Got 50 to 60 ng/microliter of DNA by Qubit dsDNA HS assay protocol.

Additional Notes

None

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Summary

The Good

Simple and easy perform assay.

The Bad

If you leave Qubit dsDNA HS fluorescent dye unprotected from light, it will bleach and loose its fluorescence.

The Bottom Line

This kit is highly recommended for accurate quantification of DNA samples needed high throughput experiments.

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