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KAPA SYBR® FAST qPCR Kits |
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KAPA SYBR® FAST qPCR MasterMix
vs
ABI Power SYBR® Green Master Mix
Customer testimonial received from Anachem Ltd.
The performance of KAPA SYBR® FAST 2x qPCR MasterMix (ABI Prism™) and ABI Power SYBR® Green Master Mix was compared in a gene expression study measuring the effect of DHT exposure (10 nM for 16 hours) on the SMIT1 gene in prostate stromal cells. Two RNA/cDNA samples were tested with both master mix reagents, with two different primer sets, SMIT1 and GAPD (housekeeping gene). cDNA was prepared with an oligo dT with SuperScript II (Invitrogen). Reactions (20 μl) were performed with the ABI7000 real-time PCR instrument and contained 1x qPCR master mix and either 200 nM (KAPA SYBR FAST) or 500 nM (ABI Power SYBR) of each primer. Results are shown below. Ct values were earlier with KAPA SYBR FAST but relative expression results were similar between the two kits.
Suzanne Kennedy
Suzanne is Director of R&D at a small biotech company in California. She has a PhD in Microbiology and Immunology from Virginia Commonwealth University.
She was writing an article here about primers for QPCR experiments bitesizebio.com/2009/09/21/read-this-before-you-design-those-qpcr-primers/
Side note: I just want to add a plug here for a little known company that sells great PCR and qPCR enzyme mixes- Kapa Bioscience. We use their Kapa SYBR Fast qPCR Kit in our lab and virtually every primer set we test works well the first time. We had a primer set that was only 80% efficient with another enzyme mix that was >90% with the Kapa mix. You can use the fast cycling without having a special cycler and it achieves a higher sensitivity than any other mix we tried. I highly recommend it!
ChiP
“I tested four samples with both ABI and KAPA SYBR FAST reagents. Please find attached the summary of the qPCR results. The KAPA SYBR reagent can bring down the Ct values by 1.5-2, which is amazing. I will go ahead and use it for my future CHIP experiments. Thank you!”
- Junqi Song, Duke University
Speed, sensitivity and reproducibility
“The Demokritos Institute asked for a sample, and after testing it, they start using it at relatively high volumes. When we asked them why they were so happy with it, the reply was that the Institute has only one real time PCR instrument, and they share the time of its use. They found that with the master mix from Kapa they could do the double number of reactions at the same time (limited for each lab). So we’ve gained all 6 labs on this Institute that work with qPCR as customers for Kapa.”
– Lab Supplies, Greece
Gene expression and copy number variation
I work for the Microarray Core facility of the University of Basel, Switzerland. We lead genome-wide studies on the RNA (gene expression and alternative splicing) and the DNA (SNPs and Copy Number Variation). Real-time PCR is a key player for validating our results. We own a Corbett Research Rotor-Gene 3000A. After using various SYBR Green Kits over the past years we switched to SyBr Fast kit from Kapa Biosystems because we noticed a significant improvement in PCR efficiency, and especially in steepness of amplification curves. What I appreciate the most is the fact this kit works very nicely for the vast majority of primers we design without optimization and for various applications (Gene expression as well as DNA Copy Number). This universality makes our real-time PCR assays very fast to establish and our results more reliable.
- Philippe Demouging, Molecular Psychology, University of Basel
“Basically, I have tried the KAPA SYBR FAST and in actual fact, I have had more success with this in the past 2 days, then I have for the past 6 months at the equivalent genomic region (and 24 primer sets later!). Obviously, I will continue to order this in the future and our lab is a high volume lab for qPCR and there is also a high possibility that other people in my lab will try Kapa products given my success with the KAPA SYBR FAST!”
- PhillippaTaberlay, University of Southern California
Speed, sensitivity and reproducibility
“I am the person in charge of the testing. Here are the reports for you to review. Personally I like the Kapa kit a lot. It certainly has some advantages over the other kits.1) Stable chemistry and enzyme. All replicates are very tight in the exponential phase, even tighter than other kits at the plateau phase .2) Good sensitivity. All curves are 1-2 Ct earlier than the other kits. And the curves are still nice with low copy numbers .3) Tighter melting curve.”
- Cheng Liu, Ph.D., Application Support Technician Eppendorf North America
Optimization of standard operating procedure for quantitative
real-time PCR in equine embryos
Smits K a, Goossens K b, Van Poucke M b, Van Soom A a, Peelman L b
a Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, Merelbeke, Belgium
b Department of Nutrition, Genetics and Ethology, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, Merelbeke, Belgium
For details, please click here.
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