High-throughput-PCR screen of 15,000 transgenicPhyscomitrella plants
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TL;DR
An easy-to-handle protocol was developed for the rapid screening of a large number ofPhyscomitrella plants and about 90% of the plants could be identified as either transgenic or nontransgenic in a single PCR reaction.
Abstract
An easy-to-handle protocol was developed for the rapid screening of a large number ofPhyscomitrella plants. Genomic DNA was isolated without liquid nitrogen or mechanical disruption of plant material and subsequently screened by PCR. This protocol has been applied successfully to approximately 15,000 mutants (800–1000 plants per wk). About 90% of the plants could be identified as either transgenic or nontransgenic in a single PCR reaction. The results of the PCR-based identification were verified by an additional round of selection of mutants on antibiotic-containing medium.