Effect of DMSO on PCR of Porphyra yezoensis (Rhodophyta) gene

Effect of DMSO on PCR of Porphyra yezoensis (Rhodophyta) gene

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Journal of Applied Phycology 15: 555–557, 2003. © 2003 Kluwer Academic Publishers. Printed in the Netherlands. 555 Note Effect of DMSO on PCR of Porphyra yezoensis (Rhodophyta) gene Graduate School of Fisheries Sciences, Hokkaido University, Hakodate, Hokkaido 041-8611, Japan ( Author for correspondence; e-mail nsaga@Fsh.hokudai.ac.jp) Received 25 June 2003; revised and accepted 8 July 2003 Key words: adjuvant, DMSO, PCR, EST clone, high GC content, Porphyra yezoensis , Rhodophyta Abstract In order to improve the predictability of results of PCR with Porphyra yezoensis Ueda genes, a study was made of possible modiFcations to the basic PCR protocol. DMSO used as an adjuvant considerably increased ampliFcation efFciency and speciFcity of PCR, the optimal concentration being 5%. This protocol allowed for DNA templates with a high GC content to be ampliFed by PCR without problem. PCR technology is widely used for various kinds of molecular biological study. Many papers and manu- als discuss in detail the conditions in±uencing the quality of PCR, probably because of its unstable character. ²actors considered include the amounts of primer, dNTP and MgCl 2 concentration, PCR buf- fer (KCl) concentration and amount of template DNA and Taq DNA polymerase (Ausubel et al., 1994). Ad- juvants such as BSA (bovine serum albumin), DMSO (dimethylsulfoxide) and glycerol have been used to improve ampliFcation efFciency (higher amount of product) and speciFcity (lack of non-speciFc products) (Henegariu et al., 1997). Also, DMSO has been used to improve cycle sequencing reaction of GC-rich DNA template although the underlying mechanism was un- known (Choi et al., 1999). We have encountered practical problems in amplifying cDNA or genomic DNA templates with a high GC content in PCR of P. yezoensis
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This note was uploaded on 08/01/2009 for the course 221 22412 taught by Professor Maz during the Spring '09 term at A.T. Still University.

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Effect of DMSO on PCR of Porphyra yezoensis (Rhodophyta) gene

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