Preparation of cDNA-silica using reverse transcriptase and its DNA sequence determination.
A new method for producing macroporous silica (suitable for high-performance liquid chromatography) with covalently attached DNA is presented. The method uses (dT)18-silica as a primer, annealed to a poly(A)-RNA template, which is then transcribed using reverse transcriptase. The RNA template is eluted and single-stranded cDNA-silica is recovered. The cDNA-silica can be sequenced using the dideoxy method. These methods provide a facile method for producing cDNA-silica of demonstrable authenticity and provide a unique approach to DNA and RNA sequencing.
Jarrett
H W
HW
Department of Biochemistry, University of Tennessee, Memphis 38163, USA.
D001483:Base Sequence; D018076:DNA, Complementary / Q000737:chemistry*; D008969:Molecular Sequence Data; D012194:RNA-Directed DNA Polymerase / Q000737:chemistry*; D017422:Sequence Analysis, DNA; D012822:Silicon Dioxide / Q000737:chemistry*
D018076:DNA, Complementary; D012822:Silicon Dioxide; D012194:RNA-Directed DNA Polymerase