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.
Scientific Contributors
Quoc Hoang, WeiWei Xu, Ro
'"/>
Source:
Page: All 1 2 3 Related biology technology :1.
A New C-Terminal GST Vector for Protein Production in S. pombe2.
New Mammalian Two-Hybrid System Detects Protein-Protein Interactions3.
Antibodies for Studying NMDA Receptor Protein Expression and
Synapse-Specific Immunolabeling4.
Simple, Sensitive, and Rapid Detection of FLAG -Tagged
Fusion Proteins5.
New Yeast Cloning System for Producing Proteins with Native Amino Acid
Sequences6.
Optimized Imaging of Protein Gels Stained with Coomassie
Brilliant Blue Dye7.
High-Level Protein Expression, One-Column Purification, and FLAG
Epitope Tagging in E. coli8.
Transfection of Green Fluorescent Protein into Human Adrenalcarcinoma Cells9.
Yeast Protein Production System Features High Yields and One-Step
Purification10.
Efficient Cleavage of Fusion Proteins to Yield Native Amino Termini11.
Expression and Purification of Recombinant Proteins That Have Native Amino
Acid Sequence