Amplification of a GC-rich sequence from barley by a two-step polymerase chain reaction in glycerol |
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| Authors: | R J Henry K Oono |
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| Institution: | (1) Queensland Wheat Research Institute, PO Box 2282, 4350 Toowoomba, Qld, Australia;(2) National Institute of Agrobiological Resources, 2-1-2, Kannondai, 305 Tsukuba, Japan |
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| Abstract: | PCR amplification of GC-rich sequences may fail due to poor denaturation or secondary structures that block elongation. Successful
amplification of a 672-bp sequence encoding the barley α-amylase/subtilisin inhibitor (69% GC) was achieved in a simple two-step
procedure with the addition of 20% glycerol and a high annealing temperature. This protocol may be useful for the amplification
of other GC-rich sequences. |
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| Keywords: | PCR GC-rich sequences α -amylase inhibitors |
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