Technique / Molecular Biology / miRNA microRNA

MicroRNA cloning protocol

MicroRNA cloning protocol

MicroRNA (miRNA) cloning protocol from Dr.Victor Ambros's lab at Dartmouth medical college.

Section I RNA ligation of 3` linker.
Section II. Gel purification of (RNA-to-3’ linker) ligation product.
Section III. RNA ligation of 5’ linker (optional, “Nelson” protocol).
Section IV. Reverse transcription
Section V. PCR amplification of cDNA
Section VI. Gel purification of cDNA
SectionVII. Cloning single-insert cDNAs.
Section VIII. Cloning Concatamerized cDNAs.
Section IX. Tailing of the ends of concatamers for T-vector ligation

Last update 12-Dec-2007, Rating of 1 votes.