Diagnostic Restriction Mapping Of The Plasmid Constructs Using ZraI Restriction Enzyme

April 19, 2018

University of Birmingham
Biosciences
Research Assistant

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Company:

New England Biolabs

Product Name:

ZraI

Catalog Number:

R0659S

I have constructed a plasmid of size 6788 bp for cloning. This plasmid was transformed into E. coli DH5 alpha cells and selected on selective agar. Plasmid prepped from these cells were subjected for ZraI diagnostic restriction mapping to check whether cells have correct plasmid. Restriction digestion was expected to give two bands of 5542 and 1246 bp. 1% agarose gel analysis revealed correct restriction mapping of the plasmid prepped from E. coli cells.

Experimental Design and Results Summary

Application

Diagnostic restriction mapping and cloning of plasmid constructs

Starting Material

1 microgram of plasmid DNA

Protocol Overview

Perform restriction digestions in a 50 microliter reaction volume with 1 microgram of plasmid DNA, 1X concentration of appropriate NEB's buffer and 1 microliter of ZraI enzyme. Incubate digestion reaction at 37 degree C for 1 hour. Then perform 1% agarose gel analysis.

Tips

Use appropriate buffer system and make sure that your plasmid DNA prep is free of salt contamination.

Results Summary

Restriction digestion of my plasmid construct with ZraI gave correct DNA fragments of sizes 5542 and 1246 bp as seen in the 1% agarose gel.

DOI or PMID #

N/A

Additional Notes

None

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Summary

The Good

Simple and easy protocol to follow.

The Bad

None

The Bottom Line

Using ZraI enzyme , my plasmid constructs were confirmed according their expected restriction maps. Very satisfied using this enzyme for diagnostic restriction mapping of my plasmid used in this study.

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