A plant DNA isolation protocol suitable for polymerase chain reaction based marker-assisted breeding

D. A. Lange, S. Peñuela, R. L. Denny, J. Mudge, V. C. Concibido, J. H. Orf, N. D. Young

Research output: Contribution to journalArticlepeer-review

22 Scopus citations

Abstract

An important, but often limiting step in marker-assisted breeding is the efficient isolation of plant DNA for polymerase chain reaction (PCR) amplification. While there are many protocols for plant DNA isolation, they tend to be time consuming or difficult to use on many samples simultaneously. We have optimized an alternative approach to plant DNA isolation that immobilizes DNA on a solid matrix, followed by processing in a 96-well microtiter plate. A single person can isolate DNA and initiate PCR-based analyses for 96 individuals in one day. The DNA samples are suitable for several PCR-based procedures, including random amplified polymorphic DNA (RAPD), microsatellite (simple sequence repeats), and amplified fragment length polymorphism (AFLP) analyses. Because the DNA is immobilized on a solid matrix and processed in 96-well plates, this protocol could be modified for robotic manipulation.

Original languageEnglish (US)
Pages (from-to)217-220
Number of pages4
JournalCrop Science
Volume38
Issue number1
DOIs
StatePublished - Jan 1 1998

Fingerprint Dive into the research topics of 'A plant DNA isolation protocol suitable for polymerase chain reaction based marker-assisted breeding'. Together they form a unique fingerprint.

Cite this