Abstract
We describe a novel method for the generation of RNA probes based on the direct in vitro transcription of DNA templates amplified by polymerase chain reaction (PCR) using primers with sequence hybrids between the target gene and those of the T7 and T3 RNA polymerases promoters. This method circumvents the need for cloning and allows rapid generation of strand-specific RNA molecules that can be used for the identification of genes in hybridization experiments. We have successfully applied this method to the identification of DNA sequences by Southern blot analysis and library screening.
Original language | English (US) |
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Pages (from-to) | 113-120 |
Number of pages | 8 |
Journal | Journal of Biochemical and Biophysical Methods |
Volume | 26 |
Issue number | 2-3 |
DOIs | |
State | Published - May 1993 |
Keywords
- Hybridization
- In vitro transcription
- Kinesin
- Polymerase chain reaction
- RNA probe
ASJC Scopus subject areas
- Biophysics
- Biochemistry