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Annals of Botany 80: 697-700, 1997
© 1997 Annals of Botany Company

COMMUNICATION

In situPCR on Plant Material with Sub-cellular Resolution

BO JOHANSEN

University of Copenhagen, Botanical Institute, Botanical Laboratory, Gothersgade 140, DK-1123, Copenhagen, K, Denmark

April 9, 1997 ; July 11, 1997 .

ABSTRACT

In situPCR and reverse transcribedin situPCR have been tested on leaves of sugar cane fixed in FAA, 4% PFA or 2% PFA+2.5% GA.In situPCR amplification of the gene coding for ribulose-1,5-bisphosphate carboxylase/oxygenase (rbcL) was successfully performed following all three fixation protocols. As expected the PCR product was restricted to the plastids of all cells. Reverse transcribedin situPCR was performed onrbcL mRNA and in this case the PCR product was restricted to the plastids of the bundle sheath cells. This is the first report ofin situPCR on plant material and only the second report ofin situPCR with sub-cellular resolution.In situPCR onrbcL may prove to be a valuable positive control for futurein situPCR studies on plant material.Copyright 1997 Annals of Botany Company

In situPCR; reversed transcribedin situPCR; rbcL; C4-plants


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