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RNA, DNA, and Protein from a Single Sample

Reagent is suitable for use in real-time and end-point RT-PCR, amplification/labeling for microarray analyses, in vitro translation, cDNA synthesis, Northern blotting, and RNase protection assays. RNA preparations isolated using bromochloropropane often do not require an additional DNase treatment, but Ambion's TURBO DNA-free Kit can be used to remove residual genomic DNA before RT-PCR assays. In addition, PCR, Southern blotting, and cloning can be performed using DNA, and denaturing polyacrylamide gel electrophoresis, and Western blot assays can be performed using protein isolated with TRI Reagent. See www.ambion.com/prod/tri for more information and detailed protocols.

Figure 1. Bromochloropropane (Phase Separation Reagent) Improves Genomic DNA Removal in the TRI Reagent Single-Step RNA Isolation Method. Total RNA was isolated from RNAlater-treated mouse liver, kidney, brain, lung, heart, and spleen and from fresh cultured human cells using the standard TRI reagent protocol with either bromochloropropane (BCP) or chloroform (CHCl3).(A) Genomic DNA removal was assessed by one-step, (+) or () RT, quantitative PCR targeting mouse or human TATA binding protein (TBP) mRNA and gene sequences, and is presented as percent of residual gDNA. (B) Total RNA yield was determined by A260 using the NanoDrop spectrophotometer, and RNA yield (g/mg) was normalized to mg of mouse tissue input or to 0.5 x 105 human cultured cells.


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