Description: Protocol for the extraction of RNA from lipid tissues (≤ 10mg). This process disrupts lipid tissues with a pellet pestle in the presence of a mono-phasic solution containing chaotropic denaturants phenol and guanidine thiocyanate. The solution is homogenized using a syringe and 21 g needle, and separated into an aqueous phase and organic phase with the addition of chloroform. The RNA is recovered from the aqueous phase and purified using silica-based membrane technology with on column DNase digestion. RNA molecules < 200 nt are selectively excluded.
Description: Protocol for the extraction of RNA from < 5×10⁵ animal cells. This process harvests and disrupts human or animal cells in the presence of a mono-phasic solution containing chaotropic denaturants phenol and guanidine thiocyanate. The solution is homogenized using a syringe and 21 g needle, and separated into an aqueous phase and organic phase with the addition of chloroform. The RNA is recovered from the aqueous phase and purified using silica-based membrane technology with on column DNase digestion. RNA molecules < 200 nt are selectively excluded.
Description: Protocol for purifying total RNA with DNase treatment. This process purifies and concentrates RNA (< 45 ug) and removes contaminant DNA using silica-based membrane technology and DNase digestion. All RNA molecules < 200 nt are selectively excluded.
Description: Introduction to ethanol-precipitations of nucleic acids, with explanation of principles. Includes a protocol for the precipitation of RNA from animal cells or tissues with troubleshooting guide.
Description: Introduction to phenol-chloroform extractions with brief history and molecular theory of technique. Includes a protocol for the isolation of RNA from animal cells or tissues with troubleshooting guide.