Description
The GRS Total RNA Kit – Bacteria – provides an efficient and fast method for the purification of high quality total RNA (including mRNA, tRNA and rRNA) from Gram-positive and Gram-negative bacteria. Eluted purified RNA is suitable for RT-PCR, Northern Blotting, mRNA selection, cDNA synthesis, and primer extension.
Principle
The GRS Total RNA Kit – Bacteria – uses Lysozyme (provided) combined with a Bacteria Lysis Buffer to lyse bacterial cell walls consisting of the peptidoglycan layer efficiently. Detergents and chaotropic salts then further lyse cells and denature proteins/inactivate RNAse. Optional DNAse treatment can be included in the protocol to remove undesired trace amounts of DNA. The buffer system is pptimized to allow selective binding of RNA to the glass fiber matrix of the spin column (1). Contaminants are completely removed using a Wash Buffer (containing ethanol) in a simple centrifugation step. The purified RNA is subsequently eluted with RNAse-Free Water. The entire procedure can be completed within 20 minutes with typical RNA yields of 5 to 60 μg.