Furthermore we would like to inform you that we have print a 1 page flier about the Kapa products productoverviewv3.08_a4_distributors.pdf and you can get this flier free of charge, whenever you'll place an order..This flier is a very nice marketing tool, showing all flagship products from Kapa Biosystems as well as a brief explanation about the technology platform. In front of customers I already realized that they'll be interested as soon as they understood that Kapa Biosystems has a pretty unique technology which allows the development of unique enzymes with great benefits for PCR experiments.

That's all for today, I hope this information will be useful for you.

Keep the ball rolling

Juergen

Direct PCR from whole Blood

Mohamed Jama1 Ping Yu¹, and Elaine Lyon^1,2
1= ARUP Institute for Clinical and Experimental Pathology, Salt Lake City, Utah.
2= Department of Pathology, University of Utah, Salt Lake City, Utah.

Introduction:

The advent of PCR technology has transformed the clinical diagnostic laboratory. Typically, DNA used in PCR assays is extracted according to established procedures which are time- consuming and increase the overall cost and turn-around time of these clinical assays. Genetic testing directly from blood has not been routinely used with regular Taq DNA polymerases, due to the presence of multiple PCR inhibitors in whole blood.. To counter the inhibition of heme group, immunoglobin G (IgG), sodium citrate, heparin, or EDTA in whole blood, two main approaches have been used. KAPA Biosystems has developed a special Taq DNA Polymerase by engineering Taq that can withstand higher concentrations of blood inhibitors by selective pressure. Taq that has evolved in this environment offers the ability to amplify DNA fragments directly from blood. Others (Blood Direct™ PCR buffers and EzWay™ Direct PCR buffer) have formulated buffers with higher pH and/or with PCR additives that will stabilize Taq and protect it from inhibitors in whole blood.

Conclusion:

All samples tested on the four assays performed very well from whole blood using the 2 different buffers and the KAPA Blood PCR kit, giving 100 % concordance with results from extracted DNA. We found these two buffers and blood kit to be compatible with fresh, stored, and dried blood samples on Guthrie cards. The process requires very little human blood and was compatible with a variety of commercially available DNA polymerases, (except for AmpliTaq DNA polymerases with Blood Direct™ PCR buffer).

Of great interest was the amplification of Huntington disease samples. These samples contain a variable number of CAG repeats, which could be amplified directly with the KAPA Blood PCR Kit B. The highest repeat we tested was 18/53 but it will be interesting to see how long of a CAG repeat can be amplified. The highest CAG repeats we have been able to amplify from extracted DNA has been in the mid 90’s.

The STR analysis of 3 whole blood samples produced the expected DNA profiles. The STR profiles generated from whole blood exhibited peak height balance across all loci as expected for high quality DNA. There were no differences between the quality of DNA profiles obtained from extracted DNA and those obtained from whole blood amplified samples. The direct amplification from whole blood and Guthrie cards made this attractive for large scale newborn screening for early disease detection and intervention.

These three procedures provide alternatives to the tedious DNA purification processes and may prevent possible errors inherent to the DNA extraction procedure, such as sample switches or contamination. We found that the buffers offered a convenient way to PCR, especially on a limited amount of sample, and allowed one to test different types of Taq as the buffers are compatible with a variety of commercially available DNA polymerases. The only drawback to this method is that, when loading samples for analysis onan ABI 3100 instrument, the samples must first be spun down and only supernatant can be used for loading as other negatively- charged products would compete for injection.

kapabloodkit-utah.pdf

No information needed?

Nippon Genetics will focus on the introduction of innovative reagents and instruments for the field of Life Science Research and Diagnostics. If these product are not of interest for you, please send us a blank email and type unsubscribe in the subject field and we will not send you any further emails.