A simple and loss-free method to remove TRIzol contaminations from minute RNA samples.
Acid guanidium phenol preparations such as TRIzol allow the reproducible isolation of high-quality total RNA from various sources. However, if applied to minimal sample sizes, the quality parameters of the isolated RNA are often low. Here we present an approach to improve the 260/280- and 230/260-nm ratios of such preparations as well as the ratio of the 18S/28S RNA. A simple extraction with 1-butanol eliminates smearing of the 28 S RNA and restores the characteristic ultraviolet (UV) spectrum of highly purified RNA. Application of the method is demonstrated for fluorescence-activated cell sorting (FACS)-sorted cells where the population of interest is often small.
Krebs
Stefan
S
Laboratory for Functional Genome Analysis, Gene Center of the Ludwig Maximilian University, Munich, Germany.
Fischaleck
Marlis
M
Blum
Helmut
H
D020001:1-Butanol; D005434:Flow Cytometry / Q000379:methods; D006146:Guanidines / Q000302:isolation & purification*; D010636:Phenols / Q000302:isolation & purification*; D012313:RNA / Q000302:isolation & purification*
D006146:Guanidines; D010636:Phenols; C411644:trizol; D012313:RNA; D020001:1-Butanol