Purely RNA

Fundamental laboratory techniques such as Northern blot analysis, RNA protection assays, in situ hybridization, and reverse transcriptase-PCR (RT-PCR) require high-quality, highly purified RNA samples. Preparing such samples is often laborious at best, because RNAses-both stable and omnipresent-can easily degrade the samples. In the past, RNA work was often left to dedicated labs, with dedicated work areas, equipment, and some very meticulous workers. Fortunately, the technology continues to evo

Deborah Stull
Nov 11, 2001
Fundamental laboratory techniques such as Northern blot analysis, RNA protection assays, in situ hybridization, and reverse transcriptase-PCR (RT-PCR) require high-quality, highly purified RNA samples. Preparing such samples is often laborious at best, because RNAses-both stable and omnipresent-can easily degrade the samples. In the past, RNA work was often left to dedicated labs, with dedicated work areas, equipment, and some very meticulous workers. Fortunately, the technology continues to evolve, and numerous RNA-purification protocols now exist that allow researchers to focus on specific scientific questions instead of on sample quality. These improvements are making RNA isolation more routine, enabling labs that lack RNA-handling expertise to venture into the arena of gene expression analysis.

The first step to RNA recovery is cell lysis, which depends on the denaturation of the proteins that comprise the cellular membrane. In 1979 John Chirgwin and colleagues devised a method for disrupting cellular membranes without denaturing nucleic acids....

Interested in reading more?

Become a Member of

Receive full access to digital editions of The Scientist, as well as TS Digest, feature stories, more than 35 years of archives, and much more!
Already a member?