Mutagenesis Kit From New England Biolabs Enables Rapid, Site-Specific Mutagenesis of Double-Stranded Plasmid DNA

January 24, 2024

UNC Chapel Hill

Overall

Quality of Results

Ease-of-Optimization

What do these ratings mean?
Write a Review

Company:

New England BioLabs

Product Name:

Q5 Site-Directed Mutagenesis Kit

Catalog Number:

E0554S

The Q5 Site-Directed Mutagenesis Kit enables rapid, site-specific mutagenesis of double-stranded plasmid DNA in less than 2 hours. The kit utilizes the robust Q5 Hot Start High-Fidelity DNA Polymerase along with custom mutagenic primers to create insertions, deletions, and substitutions in a wide variety of plasmids.

Experimental Design and Results Summary

Application

Mutagenesis

Starting Material

Cultured mammalian cells

Tips

I use the dilution as recommended

Results Summary

After PCR, the amplified material is added directly to a unique Kinase-Ligase-DpnI (KLD) enzyme mix for rapid (5 minutes), room temperature circularization and template removal. Transformation into high-efficiency NEB 5-alpha Competent E. coli, provided with the kit, ensures robust results with plasmids up to at least 20 kb in length.

DOI or PMID #

N/A

Image Gallery

Summary

The Good

Works well with all cell lines.

The Bad

Can't think of anything bad.

The Bottom Line

Advantages:non-overlapping primer design ensures robust, exponential amplification, generating a high percentage of desired mutations from a wide range of templates. Intramolecular ligation and transformation into NEB high-efficiency competent cells results in high colony yield, Extremely low error rate DpnI background reduction permits a wide range of starting template concentrations.

Join the discussion