Thermostable Reverse Transcriptase

dacoo at BE.AC.ULB dacoo at BE.AC.ULB
Thu Jun 18 05:33:22 EST 1992


Hello netters,
                        Needing Help
 
        I would like to determine the 5' extremity of Rna by primer
extension. Since this RNA is not very abundant, I would like to use a
thermostable reverse transcriptase such that I could repeat
Hybr./elong./denat. cycles, in order to amplify the cDNAs products.
        Using a higher temp, I would also benefit of the weakening of RNA
secondary structures, this would give better results.
                        BUT...
        Some people says that thermostable reverse transcriptase from cetus
works in Mn buffer, and that Mn++ ions could degrade RNA ???
 
        Does some of you experienced this ?
 
        Any suggestions...
        Any comments ...
        Any help          would be very appreciated
 
 
Coornaert David
Brussel Free University
Belgium
 
*** dacoo at ulb.ac.be ***



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