This application note demonstrates a simple and rapid way to produce template DNA for PCR from small insects such as the immature stages of scale insects, many of which are difficult to distinguish morphologically. It describes the development of a species diagnostic test using prepGEM followed by multiplex PCR using sequence variation in the cytochrome oxidase genes to distinguish among three species of scale insects found on kiwifruit leaf and fruit material. This approach produces suitable amounts of DNA as a template for PCR, even from immature stage insects less than 1 mm in length, and is scalable to high throughput automation with minimal physical manipulation of the sample.
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MicroGEM’s prepGEM Universal is a widely-used, adaptable reagent kit suitable for DNA extractions from life science samples. Because the entire extraction is done in a single-tube with no washes or additional steps, there is no loss of DNA, enabling extractions from even a single cell.
Extracts DNA from blood, saliva, and tissue including animal (ear punches, mouse tails, insect) and cell culture
Produces single-stranded DNA for STR, PCR, qPCR, whole genome amplification, amplicon sequencing
Perfect for CRISPR/Cas9 gene editing with NGS
Each extraction kit contains prepGEM, Histosolv, BLUE buffer, RED+ buffer, and ORANGE+ buffer