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 two 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).
For more detailed protocol information on mouse genotyping from crude lysates using end-point PCR download the Application Note here>>
Ordering and product information for KAPA2G Robust DNA Polymerase here>>
Ordering and product information for KAPA SYBR® FAST qPCR Kits here>>