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pCMV-SPORT6 Library Method Overview

Prior to January 2001, Life Technologies, Invitrogen Corp., Carlsbad, CA, constructed non-normalized (standard) libraries in the pCMV-SPORT6 vector, which was prepared at Invitrogen. Briefly, the library method was as follows:
  • First strand cDNA was synthesized from polyA+mRNA using Invitrogen Superscript II RT and an oligo-dT primer containing a Not I site.
  • Second strand cDNA was synthesized.
  • Sal I adapters were ligated onto the double stranded cDNA which was then digested with Not I creating 5'-Sal I/3'- Not I inserts.
  • This cDNA was size-fractionated on a column and ligated into the pCMV-Sport6 vector.