Diagnostic Restriction Mapping The Plasmid Construct With AatII

May 04, 2018

University of Birmingham
Biosciences
Research Assistant

Overall

Quality of Results

Ease-of-Optimization

What do these ratings mean?
Write a Review

Company:

New England Biolabs

Product Name:

AatII

Catalog Number:

R0117S

I have constructed a plasmid called pTS30-flaB of a size 7.13 kb. The plasmid was prepped from the transformed E. coli cells and was subjected to diagnostic restriction mapping with AatII from NEB. Restriction was expected to give 3 DNA fragments of size 4.9, 1.33 and 0.9 kb size. Indeed restriction of the plasmid pTS30-flaB with AatII revealed expected restriction pattern.

Experimental Design and Results Summary

Application

Diagnostic restriction mapping and cloning of the plasmid constructs

Starting Material

1 microgram of plasmid DNA

Protocol Overview

Restriction digestion of the plasmid was performed in a 50 microliter reaction volume with 1 microgram of plasmid DNA, 10X cutsmart buffer to a final concentration of 1X and 1 microliter of enzyme. Reaction mix was incubated at 37 degree C for 2 hours and then analysed in 1% agarose gel run.

Tips

Plasmid miniprep should be of good quality without any salt contamination. Use appropriate compatible buffer for effective digestion.

Results Summary

Restriction of the plasmid pTS30-flaB with AatII revealed 3 DNA fragments of size 4.9, 1.33, and 0.9 kb. This was expected restriction pattern as seen in the 1% agarose gel run.

DOI or PMID #

N/A

Additional Notes

None

Image Gallery

Summary

The Good

Very easy and simple protocol. Not expensive enzyme to buy.

The Bad

None

The Bottom Line

Restriction mapping using AatII on my plasmid construct went well without any troubleshooting. I recommend this enzyme in cloning experiments if the plasmid construct have AatII restriction site.

Join the discussion