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:11/24/2015)... ... November 24, 2015 , ... The United States Golf Association (USGA) today ... Green Section Award. Presented annually since 1961, the USGA Green Section Award recognizes an ... turfgrass. , Clarke, of Iselin, N.J., is an extension specialist of turfgrass ...
(Date:11/24/2015)... , Nov. 24, 2015 Halozyme Therapeutics, Inc. (NASDAQ: ... in New York on Wednesday, December 2 ... Torley , president and CEO, will provide a corporate overview. ... York at 1:00 p.m. ET/10:00 a.m. PT . ... relations, will provide a corporate overview. --> th ...
(Date:11/24/2015)... Worcester, Mass. (PRWEB) , ... November 24, 2015 ... ... need to maintain healthy metabolism. But unless it is bound to proteins, copper ... Institutes of Health (NIH), researchers at Worcester Polytechnic Institute (WPI) will conduct a ...
(Date:11/24/2015)... QUEBEC CITY , Nov. 24, 2015 /PRNewswire/ - ... the request of IIROC on behalf of the Toronto ... this news release there are no corporate developments that ... price. --> --> ... --> . --> Aeterna Zentaris ...
Breaking Biology Technology:
(Date:11/18/2015)... , Nov. 18, 2015  As new scientific discoveries ... doctors and other healthcare providers face challenges in better ... and patients. In addition, as more children continue to ... patient,s adulthood and old age. John M. ... Children,s Hospital of Philadelphia (CHOP) . --> ...
(Date:11/17/2015)... 17, 2015  Vigilant Solutions announces today that Mr. ... of Directors. --> --> ... from the partnership at TPG Capital, one of the ... $140 Billion in revenue.  He founded and led TPG,s ... TPG companies, from 1997 to 2013.  In his first ...
(Date:11/12/2015)... Mass. , Nov. 12, 2015  Arxspan ... Institute of MIT and Harvard for use of ... discovery information management tools. The partnership will support ... both biological and chemical research information internally and ... will be used for managing the Institute,s electronic ...
Breaking Biology News(10 mins):