Change search
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • harvard1
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
Strategies for overcoming PCR inhibition
Show others and affiliations
2008 (English)In: Cold Spring Harbor Protocols, ISSN 1940-3402, E-ISSN 1559-6095, Vol. 3, no 3Article in journal (Refereed) Published
Abstract [en]

To design a reliable and sensitive conventional or real-time PCR method, it is crucial to select the optimal DNA polymerase(s) and/or amplification facilitator(s) for the chemicals present in the samples during amplification. PCR optimization currentlytends to focus on primer design, buffer, and thermocycling conditions to obtain specific PCR products. The amplification mixture canbe modified by the addition of PCR facilitators or by the choice of the appropriate polymerase to improve proofreading activity, PCR yield, length of amplicon, etc. Modifications can also be made to suit specific applications, such as cloning of PCR products, in vitro mutagenesis, in situ PCP, multiplex PCR, PCR ELISA, or reverse-transcription PCR, However, less effort has been devoted to overcoming the effects of PCR-inhibitory compounds. Ideally, the number of steps required to generate PCR samples should be minimized. The use of appropriate DNA polymerases and amplification facilitators, in combination with optimized sampling techniques reduces the amount of sample handling involved in the analysis. As new PCR technology develops, research inpre-PCR processing is likely to expand in response to the growing demand for rapid, robust, and simple PCR protocols. A future challenge in pre-PCR processing strategies is to design PCR protocols that integrate sampling and DNA amplification in anautomated manner. Furthermore, to monitor PCR performance in the presence of biological compounds, mathematical models are required for the objective interpretation of PCR results. Once the validation parameters of the PCR assay of interest have been studied, the specific sample matrix can be characterized with respect to VCR interference. After obtaining this information, pre-PCR processing strategies can be designed to optimize the protocol for a specific sample. Copyright © 2008 by Cold Spring Harbor Laboratory Press.

Place, publisher, year, edition, pages
2008. Vol. 3, no 3
Keywords [en]
buffer, DNA polymerase, article, enzyme linked immunosorbent assay, gene amplification, methodology, polymerase chain reaction, process optimization, quantitative analysis, real time polymerase chain reaction, reverse transcription polymerase chain reaction
National Category
Natural Sciences
Identifiers
URN: urn:nbn:se:ri:diva-39032DOI: 10.1101/pdb.top20Scopus ID: 2-s2.0-44949138609OAI: oai:DiVA.org:ri-39032DiVA, id: diva2:1324725
Available from: 2019-06-14 Created: 2019-06-14 Last updated: 2019-06-14Bibliographically approved

Open Access in DiVA

No full text in DiVA

Other links

Publisher's full textScopus

Authority records BETA

Löfström, Charlotta

Search in DiVA

By author/editor
Löfström, Charlotta
In the same journal
Cold Spring Harbor Protocols
Natural Sciences

Search outside of DiVA

GoogleGoogle Scholar

doi
urn-nbn

Altmetric score

doi
urn-nbn
Total: 1 hits
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • harvard1
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
v. 2.35.7