Features | Partner Sites | Information | LinkXpress
Sign In
GLOBETECH PUBLISHING
GLOBETECH PUBLISHING
PZ HTL SA

Robust Site-Directed Mutagenesis Kit Enables Standard and Novel Applications

By BiotechDaily International staff writers
Posted on 22 May 2013
A new kit that enables rapid site-directed mutagenesis (SDM) of plasmid DNA has been designed to include unique features that expand the range of applications, including a more effective method for introduction of long insertions and deletions. A new SDM primer design web tool that supports the kit has also been developed.

New England Biolabs (NEB; Ipswich, MA, USA) introduces the “Q5 Site-Directed Mutagenesis Kit,” capable of making standard single-base modifications as well as novel manipulations such as deletions or additions of almost any length, including for the addition of nuclear localization signals to cloned proteins. It also includes unique features for difficult-to-amplify GC-rich and repetitive sequences. "The Q5 SDM Kit makes applications possible for which you might not have thought of using SDM. For example, SDM can be a much quicker and less error-prone way to add tags, make large deletions, add or remove restriction sites, and many other sequence modifications," said John Pezza, PhD, scientist at NEB’s Applications and Product Development.

NEB adopted a nonoverlapping primer design to allow for longer modifications. The most popular SDM methods use overlapping primers that limit the length of insertions or deletions. NEB's approach enables exponential amplification and more efficient plasmid transformation, which also facilitates the successful modification of highly repetitive or GC-rich sequences. NEB has also released “NEBaseChanger,” an SDM primer design web tool that supports the Q5 SDM Kit. Developed by Sanjay Kumar, PhD, Senior Research Scientist at NEB, it includes a novel feature - the application of nearest neighbor thermodynamics for matched and mismatched bases to design mutational SDM primers and calculate custom annealing temperatures. "These adjustments increase the odds that the experiment will work. Most, if not all other design tools, are not able to take mismatches into account," said Dr. Pezza. NEBaseChanger also provides live primer design updates, so users can make iterative changes to their primers and watch how details like reading frame and Tm are affected.

The Q5 SDM Kit includes the robust Q5 Hot Start High-Fidelity Polymerase, which has been tested to amplify sequences with a fidelity 100-times greater than Taq and twice that of “Phusion” DNA Polymerase. The kit also includes high-efficiency NEB 5-alpha Competent E. coli. Robust results have been observed with plasmids up to at least 14 kb.

Related Links:

New England Biolabs
Q5 Site-Directed Mutagenesis Kit



comments powered by Disqus

Channels

Genomics/Proteomics

view channel

Researchers Discover New Data on Protein Kinase A

By employing X-rays and neutron beams, a team of researchers have gleaned new information about protein kinase A (PKA), an omnipresent master control protein that helps regulate basic cellular functions such as energy consumption and interactions with neurotransmitters, hormones, and drugs. The scientists who conducted... Read more

Drug Discovery

view channel

Molecule in Green Tea Used as Carrier for Anticancer Proteins

A molecule that is a key ingredient in green tea can be employed as a carrier for anticancer proteins, forming a stable and effective therapeutic nanocomplex. This new discovery could help to construct better drug-delivery systems. Some cancer treatments depend on medication comprising the therapeutic drug and a carrier... Read more

Business

view channel

Interest in Commercial Applications for Proteomics Continues to Grow

Increasing interest in the field of proteomics has led to a series of agreements between private proteomic companies and academic institutions as well as deals between pharmaceutical companies and novel proteomics innovator biotech companies. Proteomics is the study of the structure and function of proteins.... Read more
 
Copyright © 2000-2014 Globetech Media. All rights reserved.