1 / 13

General guidelines for primer design

General guidelines for primer design. 18-30 nucleotides G/C content: 40-60% Avoid complementary sequences of primers (especially at the 3’ end) Avoid mismatches at the 3’ end Avoid 3 or more G or C at the 3’ end Avoid a 3’ end T.

dgarceau
Download Presentation

General guidelines for primer design

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript


  1. General guidelines for primer design • 18-30 nucleotides • G/C content: 40-60% • Avoid complementary sequences of primers (especially at the 3’ end) • Avoid mismatches at the 3’ end • Avoid 3 or more G or C at the 3’ end • Avoid a 3’ end T

  2. Primer design to eliminate amplification from contaminating genomic DNA

  3. Primer design to detect amplification from contaminating genomic DNA

  4. Primer design and analysis tools • Vector NTI • Primer 3 • IDT SciTools PrimerQuest Oligo Analyzer

  5. Real-time PCR • QPCR • Real-Time Vs Traditional PCR Post PCR processing Poor agrose gel resolution (semi-quantitative) Detect end point of the reaction

  6. http://www.appliedbiosystems.com/support/tutorials/pdf/rtpcr_vs_tradpcr.pdfhttp://www.appliedbiosystems.com/support/tutorials/pdf/rtpcr_vs_tradpcr.pdf

  7. http://www.appliedbiosystems.com/support/tutorials/pdf/rtpcr_vs_tradpcr.pdfhttp://www.appliedbiosystems.com/support/tutorials/pdf/rtpcr_vs_tradpcr.pdf

  8. http://www.appliedbiosystems.com/support/tutorials/pdf/rtpcr_vs_tradpcr.pdfhttp://www.appliedbiosystems.com/support/tutorials/pdf/rtpcr_vs_tradpcr.pdf

  9. ΔCt, ΔΔCt method to detect fold change

More Related