An efficient one-step site-directed and site-saturation mutagenesis protocol

Clicks: 5
ID: 296550
2004
Free PDF
Article Quality & Performance Metrics
Overall Quality
Not rated
Combines reader engagement with the AI quality analysis. This article has not been analysed, so there is no overall score — reader engagement is measured and shown alongside.
AI Quality Assessment
Not analyzed
Readership in this journal
Steady

Ranked #1,034 of 1,217 articles by views in Nucleic Acids Research

Most read Least read

Bar heights use a square-root scale. Only the 120 most-read articles are drawn; the journal has 1,217 in total.

Mint this article as an NFT
Not yet minted

Create a permanent, verifiable on-chain record of this article on the Scimatic Network. The NFT is held in your Journament account, and you can withdraw it to your own wallet at any time.

5 SUSD one-off · no wallet required
Sign in to mint
Abstract
We have developed a new primer design method based on the QuickChange TM site-directed mutagenesis protocol, which significantly improves the PCR amplification efficiency.This design method minimizes primer dimerization and ensures the priority of primer-template annealing over primer self-pairing during the PCR.Several different multiple mutations (up to 7 bases) were successfully performed with this partial overlapping primer design in a variety of vectors ranging from 4 to 12 kb in length.In comparison, all attempts failed when using complete-overlapping primer pairs as recommended in the standard QuickChange TM protocol.Our protocol was further extended to site-saturation mutagenesis by introducing randomized codons.Our data indicated no specific sequence selection during library construction, with the randomized positions resulting in average occurrence of each base in each position.This method should be useful to facilitate the preparation of high-quality site saturation libraries.
Reference Key
openalex_W2144175569 Use this key to autocite in the manuscript while using SciMatic Manuscript Manager or Thesis Manager
Authors Lei Zheng
Journal Nucleic Acids Research
Year 2004
DOI
10.1093/nar/gnh110
URL
https://doi.org/10.1093/nar/gnh110
Keywords Keywords not found

Citations

No citations found. To add a citation, contact the admin at info@scimatic.org

Comments

No comments yet. Be the first to comment on this article.