Isolation Of Biotinylated DNA Post PCR Amplification

November 01, 2017

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:

Dynabeads™ MyOne™ Streptavidin C1

Catalog Number:

65001

I have used Dynabeads Myone Streptavidin C1 for pulling DNA fragments amplified by biotinylated primers. Use of these beads greatly enhanced yield of specific DNA fragments being enriched for my downstream processing in amplicon sequencing process. Used 1mg beads and ended with recovering 10 microgram of DNA fragments amplified with biotinylated primers.

Experimental Design and Results Summary

Application

Enrichment of DNA fragments amplified with biotinylated primers in a PCR

Starting Material

20 microgram of PCR product amplified by biotinylated primers

Protocol Overview

1. Wash dynabeads according to kit's instructions provided. I used 1mg beads per sample.2. Immobilize the DNA fragments (biotinylated) according to immobilization of nucleic acid protocol given in the kit's protocol exactly.3.To release nucleic acids/DNA fragments after immobilization, Incubate beads in 10 mM EDTA, pH 8.2 with 95% formamide at 65°C for 5 min,or 90°C for 2 min. >96% of immobilized biotinylated DNA is typically dissociated.

Tips

Avoid introducing air bubbles while mixing Dynabeads Myone Streptavidin C1.

Results Summary

Good recovery DNA fragments amplified with biotinylated PCR primers. Yield was almost 70 % and quality was also good as assessed by Nanodrop measurement (260:280 was about 1.8).

DOI or PMID #

N/A

Additional Notes

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Summary

The Good

Simple and quick protocol. Less hands on time. Can be used in automated process for high throughput.

The Bad

Slightly expensive considering current trend in nucleic acids synthesis era.

The Bottom Line

Ideal for pulling targeted nucleic acids/ DNA fragments for specific type of DNA library sequencing.

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