PCR-Free Library Preparation for Illumina NGS

March 25, 2020

New York University
Biology
Research Scientist

Overall

Quality of Results

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Company:

Bioo Scientific Corporation

Product Name:

NEXTflex PCR-Free DNA-Seq Kit

Catalog Number:

NOVA-5142-01

The NEXTflex PCR-Free DNA-Seq Kit is a step-by-step PCR free DNA library preparation kit for DNA samples ranging from 500 ng to 3 ug in starting material. The kit can be multiplexed using Revvity's PCR-free barcodes (must be purchased separately) and requires access to a non-enzymatic fragmentation method for best results.

Experimental Design and Results Summary

Application

PCR-free DNA library Preparation

Starting Material

Genomic DNA (500 ng to 3 ug of starting material required)

Tips

Read manual thoroughly before starting library preparation; Ensure all reagents and equipment are prepped and available for workflow continuity.

Results Summary

Quantitative and qualitative QC should be conducted post-library preparation with this kit to ensure high quality sequencing results are attainable; Library concentration can be determined by qPCR, and library fragment size and distribution can be determined by TapeStation or equivalent.

DOI or PMID #

N/A

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Summary

The Good

Reagent color coding (see image) makes it very easy to follow the well laid out instruction manual; Safe stopping points are well defined within the manual; Quality of results is consistently high.

The Bad

Samples with starting concentrations below 500 ng can not be used;

The Bottom Line

The kit is ideal for high concentration genomic DNA samples that do not require PCR amplification during the library preparation process. The protocol is generally easy to follow and time efficient.

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