Diagnostic Restriction Mapping Of Plasmid Constructs With SalI-HF

April 11, 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:

SalI-HF

Catalog Number:

R3138S

I routinely construct several different types of plasmids for cloning and expression purposes. I have constructed plasmid of a size 6134 bp. After transforming it into suitable E. coli cells, colonies selected on selective agar plate were used for plasmid propagation and extraction. Then plasmids were confirmed by restriction with SalI-HF enzyme which gives bands of 4188bp, 1764 bp and 182 bp in agarose gel analysis. Both plasmid preps confirmed by exhibiting expected restriction map with SalI-HF.

Experimental Design and Results Summary

Application

Diagnostic restriction mapping of plasmid constructs

Starting Material

1 microgram plasmid preps in a 50 microliter restriction reactions

Protocol Overview

Overall protocol is simple. Just follow the protocol instruction which comes with SalI-HF enzyme. Perform restriction digestion in a 50 microliter reaction volumes. Use 1 microgram of plasmid DNA and NEB's Cutsmart buffer to final concentration of 1X. Then add 1 microliter of SalI-HF enzyme. Incubate digestion mix to 1 hr at 37 degree C. Stop reaction by adding 6X DNA loading dye and perform 1% agarose gel analysis.

Tips

Avoid high salt concentration in the digestion reaction. Make sure that extracted plasmid DNA is free from any salt contamination.

Results Summary

SalI-HF restriction digestion with my plasmid constructs confirmed correct constructs by showing expected DNA fragments i.e 4188 bp, 1764 bp and 182 bp as seen in 1% agarose gel analysis.

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

Digestion reactions are time saver qualified and can be completed in 15 minutes.

The Bad

None

The Bottom Line

Satisfied with using NEB's SalI-HF restriction enzyme for quick diagnostic mapping of my plasmid constructs. I recommend using this enzyme if your plasmid construct does have SalI-HF restriction sites.

Join the discussion