Techniques / PCR, qPCR and qRT-PCR

register or log in to subscribe [?]
Techniques / PCR, qPCR and qRT-PCRsubscribe
Article

Methods for Relative Quantification of qPCR Data. Yes, There is More Than One.

As all of you probably know, methods for calculating relative gene expression from qPCR data include: a) double delta Ct (ΔΔCt) and b) that one other method. Chances are you’ve probably gotten...
Read On
Webinar

Decoding Cancer: Practical Advice for Working with Cellular Heterogeneity

In this webinar, you will learn about the process for genotyping cells—particularly tumor cells—and practical advice for analyzing the resulting data. The main points that we will cover are: ...
View Webinar
Sponsored Education

The Essential PCR Troubleshooting Checklist

Routine PCR? Let's be honest, there's no such thing. Even with the simplest PCR reaction things can go wrong, so you need to have a good checklist of ideas for PCR troubleshooting and rectifying the problem....
Read On
Article

SPUD’s Your Bud When it Comes to Sensitive qPCR

There's piloting a brand new technique for the first time. Then, there’s jumping through hoops trying to get an established lab technique to work. The former, in contrast to the latter, is expected to be...
Read On
Sponsored Education

PCR Problems? Try an Additive

You've tried all the usual stuff, and checked the primer sequences twice, but still can't get that PCR fragment amplified. It's time to enter the strange world of PCR additives. Over the years a variety of...
Read On
Article

How Thermophilic Bacteria Survive, Part II: DNA

In part I, I answered the question, "How do proteins in thermophiles survive under high temperatures?" In this part, I'll look look at how nucleic acids survive -thrive, even- in conditions that are too hot...
Read On
Article

Site-Directed Mutagenesis Tips and Tricks

There are a few different ways of approaching site-directed mutagenesis. Here, I’ll give you a quick introduction to inverse PCR and why it’s useful, as well as going through a full protocol for SDM using modified primers!
Read On
Article

Troublesome Site-Directed Mutagenesis: Troubleshooting Your Experiment for Stubborn Mutations

As is sadly the case in many experiments, site-directed mutagenesis (SDM) does not always work the way we would like it to the first time around. Here are a few tips to help you on your way when trying to troubleshoot a bothersome SDM reaction!
Read On
Article

No More White Elephants! – Consider this Before Buying a Real-time PCR Cycler

Does your lab have a closet full of white elephants; once expensive instruments that are no longer fit for purpose, or have broken down? In many cases, all of that wasted money and resource could have been...
Read On
Article

When PCR Gets RACE-y: From Unknown mRNA Segments to Sequenced cDNA

Normally you need two primers to amplify your segment of interest - one for the 3' end of your segment of interest and one for your 5' end. But if you don't know the sequence of the regions you're hoping to...
Read On