PCR Optimization
PCR Sensitivity and Specificity
Category: Optimization
Obtaining Maximum PCR Sensitivity and Specificity
from Cameron N. Gundry and Matthew D. Poulson writing in PCR Troubleshooting and Optimization: The Essential Guide:
PCR is a highly sensitive and specific technique used in molecular biology laboratories everywhere. It is able to provide near 100% sensitivity and specificity with appropriately designed assays in controlled situations. However, results do not always match this potential. The most common problems in PCR arise from overlooking basic principles in assay design and optimization. Maximum PCR performance depends on key factors which include: 1) choosing an appropriate detection system, 2) using available software for the best primer and probe design, 3) assessing sample quality and controlling inhibitors, 4) avoiding amplicon and environmental contamination, 5) optimizing for reagent quality and concentration, and 6) modifying the thermal cycling protocol for optimal sensitivity and specificity. Addressing all of these factors will aid the investigator in designing high quality PCR assays.
Further reading: PCR Troubleshooting and Optimization: The Essential Guide
from Cameron N. Gundry and Matthew D. Poulson writing in PCR Troubleshooting and Optimization: The Essential Guide:
PCR is a highly sensitive and specific technique used in molecular biology laboratories everywhere. It is able to provide near 100% sensitivity and specificity with appropriately designed assays in controlled situations. However, results do not always match this potential. The most common problems in PCR arise from overlooking basic principles in assay design and optimization. Maximum PCR performance depends on key factors which include: 1) choosing an appropriate detection system, 2) using available software for the best primer and probe design, 3) assessing sample quality and controlling inhibitors, 4) avoiding amplicon and environmental contamination, 5) optimizing for reagent quality and concentration, and 6) modifying the thermal cycling protocol for optimal sensitivity and specificity. Addressing all of these factors will aid the investigator in designing high quality PCR assays.
Further reading: PCR Troubleshooting and Optimization: The Essential Guide
Getting The Most Out of PCR
An online seminar series entitled "Getting The Most Out of PCR", is being broadcast by the popular life science blog, Bitesize Bio. Bitesize Bio is headed by Nick Oswald and Suzanne Kennedy, co-editors of the forthcoming book "PCR Troubleshooting and Optimization".
The series lineup includes many of the authors from this book and commences on May 18 with a talk from LightCycler co-inventor, Carl Wittwer, entitled "Magic in Solution: An Introduction and Brief History of PCR". This will be a great learning experience with an opportunity to ask questions and learn from experts and pioneers in the PCR field. The full program is shown below.
Click here to book your place on these excellent events.
Recommended reading: PCR publications
The series lineup includes many of the authors from this book and commences on May 18 with a talk from LightCycler co-inventor, Carl Wittwer, entitled "Magic in Solution: An Introduction and Brief History of PCR". This will be a great learning experience with an opportunity to ask questions and learn from experts and pioneers in the PCR field. The full program is shown below.
Click here to book your place on these excellent events.
- Magic in Solution: An Introduction and Brief History of PCR
Speaker: Carl Wittwer
18 May 2010 / 9am Pacific / 12pm Eastern / 5pm GMT / 6pm CET - Obtaining Maximum PCR Sensitivity and Specificity
Speaker: Cameron N. Gundry
25 May 2010 / 9am Pacific / 12pm Eastern / 5pm GMT / 6pm CET - Significance of Controls and Standard Curves in PCR
Speaker: Ian Kavanagh
01 June 2010 / 9am Pacific / 12pm Eastern / 5pm GMT / 6pm CET - The MBD2-based Enrichment Approach for Analyzing DNA methylation
Speaker: Chris Adams
08 June 2010 / 9am Pacific / 12pm Eastern / 5pm GMT / 6pm CET - The MIQE Guidelines Uncloaked
Speaker: Greg Shipley
15 June 2010 / 9am Pacific / 12pm Eastern / 5pm GMT / 6pm CET - High Resolution Melting Analysis - Beyond the SNP
Speaker: John Mackay
22 June 2010 / 9am Pacific / 12pm Eastern / 5pm GMT / 6pm CET
Recommended reading: PCR publications