geneart_mutagenesis

Site-directed mutagenesis techniques allow specific introduction of variations at desired locations within DNA, without the need for complete de novo DNA synthesis. Mutagenesis of existing genes can help you find answers for your research, such as:

  • Investigate active sites, structure-function relationships, nucleic acid–protein interactions, etc.
  • Test functional elements (promoter regions, terminator sequences, etc.)
  • Construct fusion proteins and tagged proteins
  • Design knockout constructs
  • Generate alternative splice forms
  • Conduct alanine scans to identify protein functional domains

The GeneArt™ site-directed mutagenesis service introduces single or multiple mutations (substitutions, insertions, deletions, or truncations) into existing DNA sequences, quickly and economically. We use a targeted PCR-based process to introduce genetic variation precisely where you need it, and then clone the construct into the vector of your choice. All final sequences are 100% sequence-verified and documented to help ensure that the DNA you receive is exactly what you’ve requested.

Site-directed mutagenesis service production process

Two ways to get your project started

Set up your project in our online order portal: add ”single variant“ or ”variant bundle“ to your project, and follow the instructions to enter your sequence(s).
  Download the Multiple Sequence Submission Form, enter your DNA sequence request(s) using Microsoft Excel software, and email it to our customer support team at geneartsupport@thermofisher.com.

For further information regarding this service or the order process, please contact geneartsupport@thermofisher.com.


Variants ≤4 kb can be ordered directly via the GeneArt online order portal or email.
Variants >4 kb can be requested via the GeneArt online order portal or email but will require manual review before order placement.

Variants may contain up to four of the following modifications:

  • 5'/3' deletion of any length, with up to 52 nt of new sequence on each side
  • 5'/3' modification of maximum 52 nt on each side
  • 5'/3' extension of maximum 52 nt on each side
  • Internal modifications of up to 40 nt (a maximum of 3 of these internal modification blocks are allowed)
  • Internal deletions of any length
  • Modifications (whether internal or 5'/3') have to be separated by at least 100 bp


GeneArt Mutagenesis Service (Single variants: ≤5 variants from one template)

  • 5 µg of plasmid DNA
  • Cloned in production vector; optional subcloning in customer vector available (additional costs + production time)
  • 100% sequence-verified

GeneArt Mutagenesis 6+ Service (Bundle variants: >5 variants from one template)

  • 5 µg of plasmid DNA
  • Cloned in customer-supplied vector
  • 100% sequence-verified


Single variantsBusiness days
Up to 3 kb6
>3 kb10
Bundle variantsBusiness days
<10 variants7
10–20 variants10
21–48 variants13
49–96 variants16
Additional 96 variants+5 each
For very large projects (500 variants or more)Please inquire


  • Mutations exactly where you need them, without the risk of introducing unwanted mutations
  • Unlike “quick change” strategies, the vector is not subject to PCR—just the insert
  • Insert size can be as large as 6–8 kb
  • No mutations in the vector backbone
  • Full sequence verification of insert DNA
  • Mutations can be clustered or far apart, with the same cost and production time
  • Template constructs are stored for at least 2 years at no charge, and longer-term storage is available
  • Competitive pricing and fast turnaround times
Variant criteria


Variants ≤4 kb can be ordered directly via the GeneArt online order portal or email.
Variants >4 kb can be requested via the GeneArt online order portal or email but will require manual review before order placement.

Variants may contain up to four of the following modifications:

  • 5'/3' deletion of any length, with up to 52 nt of new sequence on each side
  • 5'/3' modification of maximum 52 nt on each side
  • 5'/3' extension of maximum 52 nt on each side
  • Internal modifications of up to 40 nt (a maximum of 3 of these internal modification blocks are allowed)
  • Internal deletions of any length
  • Modifications (whether internal or 5'/3') have to be separated by at least 100 bp
Deliverables


GeneArt Mutagenesis Service (Single variants: ≤5 variants from one template)

  • 5 µg of plasmid DNA
  • Cloned in production vector; optional subcloning in customer vector available (additional costs + production time)
  • 100% sequence-verified

GeneArt Mutagenesis 6+ Service (Bundle variants: >5 variants from one template)

  • 5 µg of plasmid DNA
  • Cloned in customer-supplied vector
  • 100% sequence-verified
Processing times


Single variantsBusiness days
Up to 3 kb6
>3 kb10
Bundle variantsBusiness days
<10 variants7
10–20 variants10
21–48 variants13
49–96 variants16
Additional 96 variants+5 each
For very large projects (500 variants or more)Please inquire

Service features

  • Mutations exactly where you need them, without the risk of introducing unwanted mutations
  • Unlike “quick change” strategies, the vector is not subject to PCR—just the insert
  • Insert size can be as large as 6–8 kb
  • No mutations in the vector backbone
  • Full sequence verification of insert DNA
  • Mutations can be clustered or far apart, with the same cost and production time
  • Template constructs are stored for at least 2 years at no charge, and longer-term storage is available
  • Competitive pricing and fast turnaround times

Resources

Publications

Probing nucleosome function: A highly versatile library of synthetic histone H3 and H4 mutants

FAQs

For Research Use Only. Not for use in diagnostic procedures.

Stylesheet for Classic Wide Template adjustments