Genotyping

Genotyping refers to the process of determining genetic variations among individuals in a species. PCR-based genotyping methods have revolutionized genotyping due to significantly improved sensitivity, reduced turnaround times and simplified workflows. A plethora of PCR-based genotyping strategies – employing end-point and real-time/quantitative PCR (qPCR) – have been developed in recent years. For most genotyping laboratories higher throughput, greater efficiency (success rates, sensitivity) and shorter turnaround times remain important areas of improvement.

Kapa Biosystems offers products with significant advantages for high throughput genotyping, namely KAPA2G Robust PCR Kits (for end-point PCR protocols) and KAPA SYBR® FAST qPCR Kits (for SYBR Green® I-based qPCR protocols). Both kits contain second-generation DNA polymerases, derived from wild-type Taq through molecular evolution. KAPA2G Robust DNA Polymerase was evolved in a chemically complex environment and is designed for robust performance across a wide variety of template and assay types, whereas the KAPA SYBR® FAST DNA Polymerase was specifically engineered for fast, high efficiency qPCR in the presence of SYBR® Green I. These enzyme systems offer:

  • More robust performance and higher success rate across a wide variety of template types and amplicons, including AT- and GC-rich sequences and regions that are difficult to amplify with wild-type Taq.
  • Significantly shorter cycling times, which translate directly into faster turnaround times.
  • Higher yields and sensitivity than wild-type Taq.
  • The ability to perform genotyping from certain crude samples. These include saliva, buccal swabs and PCR-ready crude lysates prepared from mouse tail and ear clippings (in as little as 10 min).
  • Flexibility in primer design due to the ability to amplify much longer targets than wild-type Taq (particularly important for qPCR assays).
 
KAPA Mouse Genotyping Kits were designed for fast and efficient genotyping from mouse tissue in less than 1 hour. Kits contain:
  • KAPA Express Extract - 15 minute extraction using no organic solvents and requiring no pH neutralization or centrifugation.
  • KAPA2G Fast Genotyping Mix - 45 minute amplification using a novel DNA polymerase engineered for high processivity and extreme speed.
  • Available in a convenient master mix format with loading dye, in both hot start and non-hot start formulations.