- Purify total RNA directly from whole blood without prior recovery of WBCs
- Simple, fast procedure - from blood to RNA in <30 minutes
- Includes RNAlater® - store blood samples 3 days or more at room temperature
- Includes DNA-free™ Reagents to ready RNA for RT-PCR and array analysis
- Isolate and stabilize high quality, intact RNA from 0.3-0.5 ml
The RiboPure-Blood Kit isolates RNA directly from small samples of whole blood. The kit combine two methods of RNA purification to remove the contaminants that are common to blood samples (protein, heme, genomic DNA, and RNases) and that can compromise downstream procedures. The result is high yields of pure RNA directly from whole blood, eliminating the need to isolate WBCs prior to RNA purification. RNA recovered using the RiboPure-Blood Kit can be used in the most challenging applications, including real-time RT-PCR and array analysis. The RNA can subsequently be enriched for white blood cell (WBC) mRNA using one of Ambion’s GLOBINclear™ Kits to selectively remove globin transcripts.
Stabilize RNA in Whole Blood
When collecting blood samples for RNA
analysis, storage of the samples prior to RNA isolation is
often required. The RiboPure-Blood Kit contains RNAlater for
the stabilization of RNA in whole blood. The addition of RNAlater to
blood samples stabilizes the RNA and prevents its degradation.
It also "freezes" the gene expression profile of the cells.
Treated samples can be safely stored at ambient temperature
for extended periods of time (up to three days or more). As
a result, samples can be safely transported from the point
of collection to a distant lab for RNA isolation without RNA
degradation. The lower figure at right demonstrates that blood samples stored
in RNAlater yield RNA of comparable quality to blood
samples processed directly.
Simple Procedure Yields Highly Pure RNA
The RiboPure-Blood procedure consists
of three steps: 1) addition of Lysis Solution directly to RNAlater treated
or fresh whole blood, 2) initial RNA purification by phenol/chloroform
extraction, and 3) final RNA purification on a glass fiber
filter. An additional DNase treatment can then be performed
with the DNA-free Reagents included in the kit. The
entire procedure, including DNase treatment, can be completed
in less than 1 hour.
The upper figure at right shows the results of a representative
real time RT-PCR experiment in which p53, BTK, and GAPDH sequences
were amplified from the RNA isolated using the RiboPure-Blood
procedure. No genomic DNA was detectable in "no RT" control
assays.
Complete Kit for Stabilization and Isolation
of RNA
The kit provides sufficient reagents
for the stabilization and purification of RNA from 40 whole
blood samples ranging from 0.3-0.5 ml each. Average yields
of RNA are between 2-4 µg/0.5 mL of whole blood (variation
is due to differences in WBC numbers between samples). The
RNA purified with the RiboPure-Blood Kit can be used in the
most challenging applications, including real-time RT-PCR and
array analysis.
|
Alternative Product
|
Mouse RiboPure™-Blood
Recover total RNA—and optionally microRNA—directly from mouse or rat blood. Mouse RiboPure-Blood includes sample tubes pre-loaded with RNAlater to prevent RNA degradation and preserve RNA profiles. |
Trademarks, Patents, Licensing
|
