A research kit designed for use with the latest droplet digital PCR systems is able to detect simultaneously the seven most common KRAS mutations in a single ddPCR experiment.

KRAS mutations are found in 90% of pancreatic cancer cases and 30% of colorectal cancer cases, making it a gene of great interest to cancer researchers. The protein product of the normal KRAS (Kirsten rat sarcoma viral oncogene) gene performs an essential function in normal tissue signaling, and its mutation is an essential step in the development of many cancers. A single amino acid substitution is responsible for the activating mutation. The transforming protein that results is implicated in various malignancies, including lung adenocarcinoma, mucinous adenoma, ductal carcinoma of the pancreas, and colorectal carcinoma.

The new Bio-Rad Laboratories, Inc. (Hercules, CA, USA) ddPCR KRAS Screening Multiplex Kit is able to detect and quantify extremely low levels (down to 0.2%) of KRAS mutant DNA in a single assay. Bio-Rad’s ddPCR technology provides an absolute measure of target DNA molecules without the need for a standard curve. This technology enables detection of a single mutant molecule in a background of 2,000 wild-type molecules (0.05% mutation frequency).

The ddPCR KRAS Screening Multiplex Kit is compatible with all of Bio-Rad’s Droplet Digital PCR Systems, including the new AutoDG instrument. AutoDG creates droplets quickly, reproducibly, and reliably. At maximum capacity, the system can generate droplets for 96 wells in less than 45 minutes. The instrument can generate droplets for fluorescent probe-based or EvaGreen dye-based detection. The AutoDG Instrument has its own hood and HEPA filter, reducing contamination so the system can be used on a standard laboratory bench, without the need for a PCR cabinet or cleanroom.

“The KRAS Screening Multiplex Kit delivers a simple yes/no answer as to whether a KRAS mutation exists in smaller samples,” said Dr. Filip Janku of the University of Texas MD Anderson Cancer Center (Houston, USA). “If researchers were to test for the presence of each possible mutation they could end up performing seven to ten separate PCR experiments only to discover the sample was negative. Multiplex kits such as the KRAS Screening Multiplex Kit allow researchers to screen for multiple mutations in a single reaction, saving time and costly resources.”

Related Links:
Bio-Rad Laboratories, Inc.
University of Texas MD Anderson Cancer Center