A common problem in current RNA isolation methods is the contamination of RNA with polysaccharides and proteoglycans which will adversely affect migration of RNA in gel electrophoresis and inhibit the reverse transcription -polymerase chain reaction (RT-PCR). T5595-4601 is a ready to use reagent for decontamination of polysaccharides and proteoglycans from RNA preparations. Our testing shows that it inceases the purity of total RNA.
T5595-4601 can be used for RNA purification in conjunction with any RNA phenol extraction method.
We strongly recommend using T5595-2839K (Tissue, mRNA Kit for RNA Isolation, BioAssay™) for RNA isolation in combination with T5595-4601 for RNA purification.
Storage and Stability:Store at room temperature. Stable for one year from the date of receipt under proper storage condition.
Protocol:Method I. If using using T5595-2839K (Tissue, mRNA Kit for RNA Isolation, BioAssay™) for RNA isolation:(a) Mix equal volume of RNAmate with supernatant from RNAexol I and RNAexol II and store at -20°C for 30 minutes
(b) Spin at 12,000 rpm for 15 minutes at 4°C, then wash pellet with 75% ethanol. Air dry pellet and dissolve RNA pellet in DEPC water or other appropriate buffer.
Method II. If using other RNA phenol extraction reagents (e.g., TRIzol, RNAzol, etc.):(a) Mix equal volume of T5595-4601 with supernatant (upper aqueous phase) from phenol extraction, then repeat step 2 in Method I.
Note: T5595-4601 be used for precipitating RNA dissolved in DEPC water or other RNA storage solution, but will result a loss of RNA quantity.
