Perfect for PCR Product Sequencing

October 16, 2015

Loyola University Chicago
Microbiology and Immunology
Postdoc fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

Affymetrix USB

Product Name:

ExoSAP-IT® PCR Product Cleanup

Catalog Number:

78200

The goal of my project is to identify the mutation in viral genome that occurs during positive selection. Since I have tons of samples that need to be PCR amplified and sequenced, I need some reagents that will facilitate this process. The product I reviewed greatly save lots of reagents and my time.

Experimental Design and Results Summary

Application

PCR cleanup and sequencing

Starting Material

PCR product

Protocol Overview

Mix 5ul PCR product with 2ul ExoSAP-IT reagent and incubate at 37degrees for 15min, then at 80 degree for 15min. Then the final product is ready for sequencing.

Tips

Do not use it for cloning purpose

Results Summary

You will find there is no primer dimer when it runs in the agarose gel (Fig A) and the sequencing reads are good as column purification (Fig B).

Additional Notes

None

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Summary

The Good

Saves PCR reagents, purification columns, and time!

The Bad

Make sure the PCR product is what you want

The Bottom Line

Typically PCR products for sequencing need to be column purified with enough volume. This product will directly clean up the PCR primers and dNTPs from PCR products and you just need 5ul cleaned product for sequencing (Fig B). Saves PCR reagents, PCR purification kit, and time! I recommend this product for PCR product sequencing, especially high throughput.

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