Posts Tagged ‘Real-Time PCR’

High Resolution Melt Analysis Applicatons

 :: Posted by American Biotechnologist on 12-29-2009

Genomics is rapidly expanding beyond merely identifying the genetic makeup of organisms to encompass studies of how variations in genotype impact physiological functions. See the article below which was published in the October 15th edition of Genetic Engineering and Biotechnology news to learn more about high-res met analysis as an alternative to probe-based genotyping.
High-Resolution Melt Analysis Applications

Are you using the right reference genes?

 :: Posted by American Biotechnologist on 12-10-2009

Considering Real-Time PCR for gene expression analysis. Have you tested multiple reference genes or are you just going to run with your favorite such ast 18S? Check out this important article before you move forward. It could save you lots of grief in the long run.

Suitable reference genes for real-time PCR in human HBV-related hepatocellular carcinoma with different clinical prognoses
Li-Yun Fu et al. BMC Cancer 2009, 9:49

Housekeeping genes are routinely used as endogenous references to account for experimental differences in gene expression assays. However, recent reports show that they could be de-regulated in different diseases, model animals, or even under varied experimental conditions, which may lead to unreliable results and consequently misinterpretations. This study focused on the selection of suitable reference genes for quantitative PCR in human hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC) with different clinical outcomes.

Suitable Reference Gene Selection 2009

Real-Time Quantitative PCR Data Analysis Tutorial

 :: Posted by American Biotechnologist on 11-12-2009

Bio-Rad Laboratories has put together a great tutorial on QPCR analysis including a thorough review of what the graphs mean and an overview of Absolute and Relative methods of quantification. The tutorial also includes a great explanation of the differences between Livak, delta CT and the Pfaffl methods of analyzing your results.

Watch part II: normalization against a reference gene

Seminar on New Standards in Real-Time PCR

 :: Posted by American Biotechnologist on 10-27-2009

There has been a lot of buzz lately on the newly published guidelines detailing the Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE) guidelines published by Bustin et al. in Clinical Chemistry 55:4, 2009. The paper was coauthored by several important PCR authorities including Jo Vandesompele and is therefore sure to become an important standard for real-time PCR publications. Dr. Sean Taylor of Bio-Rad has put together several short youtube videos explaining the importance of MIQE and how it can be applied to the bench.

Here are the links:

Part 1 of 4

Part 2 of 4

Part 3 of 4

Part 4 of 4

Real-Time PCR Applications Guide

 :: Posted by American Biotechnologist on 10-16-2009

Nucleic acid amplification and detection are among the most valuable techniques used in biological research today. Scientists in all areas of research — basic science, biotechnology, medicine, forensic science, diagnostics, and more — rely on these methods for a wide range of applications. For some applications,
qualitative nucleic acid detection is sufficient. Other applications, however, demand a quantitative analysis. Choosing the best method for your application requires a broad knowledge of available technology.

This guide from Bio-Rad provides an introduction to many of the technical aspects of real-time PCR. It includes guidelines for designing the best real-time PCR assay for your experiments and explains how real-time PCR data are used in various applications.

To download the Guide click here (it will take a moment): Real-Time PCR Applications Guide