SIMPLE METHOD FOR RNA PREPARATION FROM CYANOBACTERIA¹

Abstract

A simple and rapid method is presented for the preparation of RNA from various cyanobacteria. Unlike other methods that require a lysis solution, lysozymes, or proteinase K, the proposed method, called the bead-phenol-chloroform (BPC) method, uses silica/zirconia beads, phenol, and chloroform to break the cells and extract RNA more efficiently. Experiments confirm that the BPC method can successfully isolate total RNA from various cyanobacterial strains without DNA contamination, and the extracted RNA samples have a relatively high purity, concentration, and yield. Furthermore, the BPC method is more rapid, simple, and economical when compared with previously reported methods.

Illumination stimulates cAMP receptor protein-dependent transcriptional activation from regulatory regions containing class I and class II promoter elements in Synechocystis sp. PCC 6803

“…RNA was isolated as previously described (Kim et al, 2006) and visualized for integrity in a formaldehyde gel. Genomic DNA was removed by two rounds of RQ1 DNase (Promega) digestion and Zymoclean (Zymo Research) column purification according to the manufacturers' instructions.…”

A comprehensive study on algal–bacterial communities shift during thiocyanate degradation in a microalga-mediated process

“…Unwanted cell debris or residual compounds were removed by repeated addition of deionized water and centrifugation, and each pellet was finally re-suspended in 500 lL of deionized water. Genomic DNA was extracted by the bead-phenol-chloroform (BPC) method (Kim et al, 2006). Briefly, each suspension was mixed with zirconia/silica beads (0.5 g of 0.5 mm diameter and 0.5 g of 0.1 mm diameter; Biospec, OK, USA) and 500 lL of saturated phenol (pH 4.3), and the mixture was beadbeaten for 60 s. The sample was then centrifuged at 14,000g for 3 min, and 490 lL of supernatant was transferred to a fresh micro-tube.…”

Morphology and phylogeny of a non-toxic invasive Cylindrospermopsis raciborskii from a Mediterranean Lake

“…culture was pelleted, glass beads of 100 and 500 lm in size (Sigma # G-4649 and G-8772) (Kim et al, 2006) were added to the tube and vortexed for 1 min in resuspension buffer. Cells were lysed by the alkaline cell lysis protocol following isopropanol precipitation and resuspended in Milli-Q water (Sambrook et al, 1982).…”