Navigation Links
Total RNA Isolation From 500 l Body Fluid

AquaPure RNA Isolation Kit
Catalog #732-6370

Cell Lysis
1. Vigorously add 2.5 ml cell lysis solution to an Oak Ridge centrifuge tube (or other tube rated for high speed) containing 500 l body fluid (e.g., cerebrospinal fluid, plasma, saliva, serum, sputum, synovial fluid, urine, whole blood).

2. Vortex for 5 sec to mix thoroughly.

3. Heat sample to 65C for 5 min to complete lysis. Cool sample to room temperature. (This step is optional when testing for certain viruses such as hepatitis C.)

Protein-DNA Precipitation
1. Add 1 ml protein-DNA precipitation solution to the lysate.

2. Cap tube and invert gently 10 times and place tube into an ice bath for 10 min. 3. Centrifuge at 15,000 x g for 5 min. The precipitated proteins and DNA will form a tight pellet.

RNA Precipitation
1. Pour the supernatant containing the RNA (leaving behind the precipitated protein-DNA pellet) into a clean Oak Ridge centrifuge tube (or other tube rated for high speed) containing 3 ml 100% isopropanol (2-propanol). If RNA yields are expected to be low, add a glycogen solution as a carrier to the isopropanol (5 l of 20 mg/ml glycogen per 3 ml isopropanol).

2. Mix the sample by capping and inverting gently 50 times.

3. Centrifuge at 15,000 x g for 5 min; depending on yield, the RNA may or may not be visible as a small, translucent pellet.

4. Pour off supernatant and drain tube briefly on clean absorbent paper. Add 3 ml 70% ethanol, cap the tube, and invert it several times to wash the RNA pellet.

5. Centrifuge at 15,000 x g for 2 min. Carefully pour off the et hanol.

6. Invert and drain the tube on clean absorbent paper and allow sample to air-dry for 15 min.

RNA Hydration
1. Add 100 l RNA hydration solution (100 l will give a concentration of 100 g/ml if the total yield is 10 g RNA).

2. Allow RNA to rehydrate for at least 30 min on ice. Alternatively, store purified RNA sample at -70 to -80C until use.

3. Before use, vortex sample vigorously for 5 sec and pulsespin. Pipet sample up and down several times to mix the sample further.

back to top


Page: All 1 2

Related biology technology :

1. New Kit Generates Exceptionally Pure Total RNA for RT-PCR
2. High-Throughput Isolation of Total RNA
3. Isolate and Analyze Total RNA from Cells Harvested by Laser Capture Microdissection
4. Fast Isolation of Total RNA from Small Numbers of Cells
5. Total RNA Isolation Protocol 1,2,3
6. Isolate It All: siRNA miRNA Total RNA Native Protein
7. Isolate Total RNA and Protein From the Same Sample
8. Isolate High Quality Total RNA from LCM Samples Suitable for Microarray and qRT-PCR Analysis
9. Recover High Yields of Total Nucleic Acid from Formalin-fixed, Paraffin-embedded (FFPE) Tissue
10. Total RNA from Whole Blood for Expression Profiling
11. RNAPrep - Total RNA Isolation Purification
Post Your Comments:

(Date:11/26/2015)... , England , November 26, 2015 ... Lightpoint Medical, an innovative medical device company specializing in imaging ... grant from the European Commission as part of the Horizon ... the company to carry out a large-scale clinical trial in ... -->      (Logo: , --> ...
(Date:11/25/2015)... , November 25, 2015 2 ... première fois les différences entre les souches bactériennes ... celles des êtres humains . Ces recherches ... et envisager la prise en charge efficace de ... diagnostiqués chez les chats .    --> ...
(Date:11/25/2015)... 25, 2015  Neurocrine Biosciences, Inc. (Nasdaq: NBIX ... and CEO of Neurocrine Biosciences, will be presenting at ... New York . .   ... approximately 5 minutes prior to the presentation to download ... presentation will be available on the website approximately one ...
(Date:11/25/2015)... , Nov. 25, 2015 Orexigen® Therapeutics, Inc. ... participate in a fireside chat discussion at the Piper ... York . The discussion is scheduled for Wednesday, ... .  A replay will be available for ... Stilwell  , Julie NormartVP, Corporate Communications and Business ...
Breaking Biology Technology:
(Date:11/10/2015)... 10, 2015 About signature ... helps to identify and verify the identity of ... as the secure and accurate method of authentication ... particular individual because each individual,s signature is highly ... when dynamic signature of an individual is compared ...
(Date:11/4/2015)... New York , November 4, 2015 ... to a new market report published by Transparency Market ... Share, Growth, Trends and Forecast 2015 - 2022", the global ... of US$ 30.3 bn by 2022. The market is ... the forecast period from 2015 to 2022. Rising security ...
(Date:10/29/2015)... 29, 2015  The J. Craig Venter Institute (JCVI) ... Synthesis and Biosecurity: Lessons Learned and Options for the ... and Human Services guidance for synthetic biology providers has ... --> --> Synthetic biology ... potential to pose unique biosecurity threats. It now is ...
Breaking Biology News(10 mins):