[Automated-sequencing] Re: Automated Sequencing of mRNA/cDNA

Alejandro Gonzalez agonzalez at gemabiotech.com
Wed Aug 17 08:03:27 EST 2005


Hi Phillips.
Thanks a lot for your info, yes I agree with you.
By away we may change doing Sanger Sequencing method using SuperScript RnaseH- Reverse Transcriptase (Invitrogen), how I have to do? we need the ddNTPs* and dNTPs and what more?, and we'd like to sequence from the mRNA. Do you have any protocol? can we make by RT-direct Sequencing?
Our RNA is total RNA from CHO cells.

Thnks

Alex G.
  ----- Original Message ----- 
  From: Alejandro Gonzalez 
  To: autoseq at net.bio.net 
  Sent: Tuesday, August 16, 2005 11:19 AM
  Subject: Automated Sequencing of mRNA/cDNA


  Hi everybody.
  I'm working in automating sequencing of the plasmid and PCR templates, using MegaBACE 1000 DNA System Sequencer and the DyEnamic TM ET dye terminator Kit (GE-ex Amersham B.). I'd like to sequence by Sanger method a mRNA template or its cDNA, if anybody know how to do? or any tips to sequence this kind of template?, if anybody could send me a protocol to prepare this sequencing reaction and if I can use same kit (of GE-Amersham) I have been using.

  Thanks a lot
  Sincerelly

  Alex G.
  Gema Biotech sa
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