Navigation Links
Total RNA Isolation: 100 l Viral Sample From Cultured-Cell Supernatant Containing Virus

AquaPure RNA Isolation Kit
Catalog #732-6370
Yield: Dependent on sample and RNA expression levels

Sample Collection and Handling
Samples may be fresh or frozen. Collect fresh virus and isolate RNA as quickly as possible; store at 18C for not more than 24 hr. For long-term storage, store at -70 to -80C.

Cell Lysis
1. Add 500 l cell lysis solution to a 1.5 ml microfuge tube that contains 100 l virus from cultured-cell supernatant.

2. Cap the tube and 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 200 l protein-DNA precipitation solution to the viral cell lysate.

2. Invert capped tube gently 10 times and place tube into an ice bath for 5 min.

3. Centrifuge at 13,00016,000 x g for 3 min. The precipitated proteins and DNA will form a tight pellet.

RNA Precipitation
1. Leaving behind the precipitated protein-DNA pellet, pour the supernatant containing the RNA into a clean 1.5 ml microfuge tube containing 600 l 100% isopropanol (2-propanol). If RNA yield is expected to be low (<1 mg), add glycogen as a carrier to the isopropanol. We recommend adding 1 l of glycogen solution (20 mg/ml) per 600 l isopropanol.

2. Cap the tube and mix the sample by inverting gently 50 times.

3. Centrifuge at 13,00016,000 x g for 3 min; depending on yield, the RNA may or may not be visible as a small, translucent pellet.

4. Pour off supernatant and drai n tube on clean absorbent paper. Add 600 l 70% ethanol and invert the capped tube several times to wash the RNA pellet.

5. Centrifuge at 13,00016,000 x g for 1 min. Carefully pour off the ethanol.

6. Invert and drain the tube on clean absorbent paper and allow to air-dry 1015 min.

RNA Hydration
1. Add 20 l RNA hydration solution (20 l will give a concentration of 50 g/ml if the total yield is 1 g RNA).

2. Allow RNA to rehydrate on ice for at least 30 min. 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 ensure adequate mixing.

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:10/9/2015)... Oct. 9, 2015  Pulmatrix, Inc., (NASDAQ: PULM ) ... investor conferences. th Annual BIO Investor ... (2:00 pm EDT). --> th Annual ... am PDT (2:00 pm EDT). --> Pulmatrix ... Growth Stock Conference on Thursday, October 15, 2015 at 8:30 ...
(Date:10/8/2015)... 2015  Sorrento Therapeutics, Inc. (NASDAQ:  SRNE; Sorrento), ... cancer and associated pain, announced today that Dr. Henry Ji, ... at the Aegis Capital Corporation 2015 Growth Conference ... Corporation 2015 Growth ConferenceDate:Friday, October 9, 2015Time:10:00 to ... The Wynn in Las Vegas, NV , ...
(Date:10/8/2015)... , October 8, 2015 ... .   --> Goldman Small Cap ... the small cap and microcap sectors, announced today ... report on PharmaCyte Biotech, Inc. (OTCQB - PMCB), ... on developing and preparing treatments for cancer and ...
(Date:10/8/2015)... 8, 2015 /PRNewswire/ - RepliCel Life Sciences Inc. (OTCQB: REPCF) ... on the development of autologous cell therapies, today announced its ... taking place in the month of October: ... RepliCel will be presenting at the annual Partnering ... Mesa to be held October 7-9 in ...
Breaking Biology Technology:
... /PRNewswire-FirstCall/ - YM BioSciences Inc. (NYSE YMI, TSX:YM), today announced that ... units at an issue price of US $1.20 for gross ... one common share and one half common share purchase warrant. Each whole ... until March 10, 2015 . , ...
... March 10 Bone Biologics, Inc. announced today ... shares from the Musculoskeletal Transplant Foundation (MTF). MTF now ... financing, MTF also provided a line of credit to ... by MTF that have allowed Bone Biologics, Inc. to ...
... ... ... ... ...
Cached Biology Technology:
(Date:9/28/2015)... 28, 2015 CLEAR, the leading ... expedited traveler service is coming to Austin-Bergstrom ... bringing a frictionless experience, serious speed and ... "CLEAR offers our travelers an expedited security ... Jim Smith , Executive Director, Austin-Bergstrom ...
(Date:9/28/2015)... JOSE, Calif. , Sept. 28, 2015 ... of human interface solutions, today announced that Lenovo has ... touch fingerprint sensor, FS4202, for its latest smartphone, the ... secure, password-free access to unlock the device and provide ... The feature-rich Natural ID FS4202 sensor ...
(Date:9/10/2015)... This report provides detailed descriptions of the sensor types ... types that will dominate in the future. Many product ... technology hype curve in the last five years before ... all of them is the prominence of sensor options ... Sensors collect data about the physical and chemical properties ...
Breaking Biology News(10 mins):
... Society today commends President Obama,s decision to sign a ... scientific evidence is used in the development of solutions ... memorandum rightly places science at the forefront of informing ... of solutions to issues such as enhancing energy security, ...
... of proteins, the molecular machines that drive all life, also ... study, in the journal Structure , reveals that after ... of innovation. The active regions of many proteins, called domains, ... host of structures that had never been seen before. This ...
... to treat critical limb ischemia in peripheral arterial ... thwarted by restenosis (the re-narrowing of the artery ... need for repeat treatments and the continued progression ... and amputation. Interventional radiologists have been studying a ...
Cached Biology News:
ma. plate stack height 70mm...
Rotor for four tube-loaded buckets; ma RCF 2550g; sold without buckets...
CQCS kit for etra KR 4i , (other units)....
A compact, high-speed single tube reader unit designed for benchtop 2D barcode reading. The unit can be used with all of ABgene's 2D barcoded tubes, including the 2ml CryoCode tube....
Biology Products: