8 Essential papers and reference guides for qPCR and more from the PCR & Real-time PCR channel at Bitesize Bio

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8 Essential papers and reference guides for Quantitative PCR (qPCR)

http://bitesizebio.com/19056/8-essential-papers-and-reference-guides-for-quantitative-pcr-qpcr/

What is PCR? – The beginner’s guide

http://bitesizebio.com/19132/pcr-basics-what-is-pcr/

qPCR Troubleshooting

http://blog.biosearchtech.com/TheBiosearchTechBlog/bid/63622/qPCR-Troubleshooting

A primer on Probe-based SNP genotyping

http://bitesizebio.com/11200/a-primer-on-probe-based-snp-genotyping/

How to scale up a PCR reaction

http://bitesizebio.com/11081/how-to-scale-up-a-pcr-reaction/

A quick tour around probe-based multiplexing qPCR

http://bitesizebio.com/11203/a-quick-tour-around-probe-based-multiplexing-qpcr/

PCR is 30! Here’s How it has Rocked the World

http://bitesizebio.com/11004/pcr-is-30-heres-how-it-has-rocked-the-world/

Q-PCR Tips and Tricks: Sobering Information and Helpful Hints for qPCR

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Dear All,

There are two recently published papers that bring a sobering perspective on the potential pitfalls and potentially problematic data published from qPCR. The first published in Nature Methods surveyed 1700 articles using qPCR with the principal conclusions that inadequate technical detail is provided to assess data quality with RNA integrity and reference gene normalization highlighted as inadequate ( http://www.nature.com/nmeth/journal/v10/n11/full/nmeth.2697.html ). The second published in Molecular Oncology surveyed 179 colorectal cancer publications for adequate normalization of qPCR data showing just three percent could be adequately assessed with common errors described

http://www.moloncol.org/article/S1574-7891(13)00188-9/

Producing high quality data from qPCR experiments is a simple matter of following a well defined procedure with some initial validation processes to assure solid results. Our recently published article entitled “The State of RT-Quantitative PCR: Firsthand Observations of Implementation of Minimum Information for the Publication of Quantitative Real-Time PCR Experiments” in the Journal of Molecular Microbiology and Biotechnology nicely summarizes this entire process in about five and a half pages. Based on the high demand for reprints, the Bio-Rad global marketing has generously purchased the Open Access licence so the paper can now be downloaded free-of-charge from the journal web site  ( http://www.karger.com/Article/PDF/356189 ). We have also recorded an accompanying, 24-minute video to guide you through the entire q-PCR process:

We continue to make every effort to summarize qPCR experimental design to help assure that you generate high quality data in a very short time frame and hope you find these tools useful. As always, your support and commitment to Bio-Rad help us in our ongoing efforts to bring you the tools and training to maximize success in your research. Feel free to contact your local sales team (cc’d on this message) to learn about our great solutions for your projects. Have a great start to the year and stay warm!!

Best,

Sean Taylor
Field Application Specialist
E:     sean_taylor@bio-rad.com

For All Technical Support
T:    1-800-424-6723
E:    tech_canada@bio-rad.com

Quantitative Western Blotting Tips: Hand Cast Stain Free Gels for Normalization of Western Blots to Produce Solid, Quantitative Data by Sean Taylor

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Dear All,

I am pleased to attach information about our TGX™ and TGX Stain-Free™ FastCast™ Acrylamide Kit which will permit you to cast your own TGX and TGX Stain Free gels. An associated video about the technology is also available (http://youtu.be/z2LKUYKKbRo). The major advantage of the Stain-Free technology for western blotting is the ability to accurately normalize by total lane density of the transferred protein as described in the attached article “A Defined Methodology for Reliable Quantification of Western Blot Data” published in the journal Molecular Biotechnology (http://link.springer.com/article/10.1007/s12033-013-9672-6). The associated video that will walk you through the article can be accessed on YouTube (http://www.americanbiotechnologist.com/blog/video-tutorial-quantitative-western-blotting/). For many western blotting applications, this means the elimination of housekeeping proteins as loading controls which cost additional time, funds and can be skew your data if your samples are not diluted appropriately to avoid signal saturation. Please read the article or watch the video for all the details.

I hope you find the Stain Free technology useful and please do not hesitate to E-mail if you have any further questions. I have cc’d your local sales specialists on this message and please feel free to consult with them regarding this great solution for achieving well normalized, quantitative western blots.

Quanti-1- 10033918_TGX_SFX FastCast Acrylamide Kit IM

Quanti-2- A Defined Methodology for Reliable Quantification of Western Blot Data_2013

Best,

Sean Taylor
Field Application Specialist
E:     sean_taylor@bio-rad.com

For All Technical Support
T:    1-800-424-6723
E:    tech_canada@bio-rad.com

How to check primers?

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How to check primers?

Just copy and paste the forward and reverse primers in these sites and choose genome(for Chip primers) or genes (for primers of mRNA or cDNA ):

1- UCSC In-Silico PCR

http://genome.ucsc.edu/cgi-bin/hgPcr

2- MFEprimer-2.0

http://biocompute.bmi.ac.cn/CZlab/MFEprimer-2.0/

3- Primer-BLAST

http://www.ncbi.nlm.nih.gov/tools/primer-blast/

4- GenomeTester

http://bioinfo.ut.ee/genometester/

LASAGNA-Search: an integrated web tool for transcription factor binding site search and visualization

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http://biogrid-head.engr.uconn.edu/lasagna_search/

 

http://www.biotechniques.com/BiotechniquesJournal/2013/March/LASAGNA-Search-an-integrated-web-tool-for-transcription-factor-binding-site-search-and-visualization/biotechniques-341032.html