compartmentalized self-replication under fast pcr cycling conditions yields taq dna polymerase mutants with increased dna-binding affinity and blood resistance
Clicks: 213
ID: 217438
2014
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.
Reader Engagement
Popular Article
30.0
/100
213 views
29 readers
AI Quality Assessment
Not analyzed
Readership in this journal
PopularRanked #714 of 875 articles by views in journal of magnetic resonance (san diego, calif : 1997)
Most read
Least read
Bar heights use a square-root scale. Only the 120 most-read articles are drawn; the journal has 875 in total.
Mint this article as an NFT
Not yet mintedCreate 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
Abstract
Faster-cycling PCR formulations, protocols, and instruments have been developed to address the need for increased throughput and shorter turn-around times for PCR-based assays. Although run times can be cut by up to 50%, shorter cycle times have been correlated with lower detection sensitivity and increased variability. To address these concerns, we applied Compartmentalized Self Replication (CSR) to evolve faster-cycling mutants of Taq DNA polymerase. After five rounds of selection using progressively shorter PCR extension times, individual mutations identified in the fastest-cycling clones were randomly combined using ligation-based multi-site mutagenesis. The best-performing combinatorial mutants exhibit 35- to 90-fold higher affinity (lower Kd ) for primed template and a moderate (2-fold) increase in extension rate compared to wild-type Taq. Further characterization revealed that CSR-selected mutations provide increased resistance to inhibitors, and most notably, enable direct amplification from up to 65% whole blood. We discuss the contribution of individual mutations to fast-cycling and blood-resistant phenotypes.
| Reference Key |
earezi2014frontierscompartmentalized
Use this key to autocite in the manuscript while using
SciMatic Manuscript Manager or Thesis Manager
|
|---|---|
| Authors | ;Bahram eArezi;Nancy eMckinney;Connie eHansen;Michelle eCayouette;Jeffrey eFox;Keith eChen;Jennifer eLapira;Sarah eHamilton;Holly eHogrefe |
| Journal | journal of magnetic resonance (san diego, calif : 1997) |
| Year | 2014 |
| DOI |
10.3389/fmicb.2014.00408
|
| URL | |
| 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.