Navigation Links
Total RNA Isolation From 50100 mg Plant Tissue


AquaPure RNA Isolation Kit
Catalog #732-6370
Expected yield range: 50200 g RNA


Method
Cell Lysis
1.Add 50100 mg (0.050.1 g) frozen ground tissue or fresh tissue to a tissue grinder tube on ice and then add 3 ml cell lysis solution. Grind frozen tissue finely in liquid nitrogen with a porcelain mortar and pestle. Keep tissue frozen until added to the tissue grinder tube.

2 . Homogenize quickly using 510 strokes with a tube pestle. Transfer sample to an Oak Ridge centrifuge tube (or other tube rated for high speed).

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

2.Invert tube 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).

2. Cap the tube. Mix the sample by inverting gently 50 times.

3. Centrifuge at 15,000 x g for 5 min; the RNA will be visible as a small, translucent pellet.

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

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

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

RNA Hydration
1. Add 300 l RNA hydration solu tion (300 l will give a concentration of 500 g/ml if the total yield is 150 g RNA).

2. Allow RNA to rehydrate on ice at least 30 min. Vortexvigorously for 5 sec, pulse-spin and carefully transfer sample to a 1.5 ml microfuge tube. Store purified RNA sample at - 7 0 C t o -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
'"/>

Source:


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:
*Name:
*Comment:
*Email:


(Date:6/23/2016)...   Boston Biomedical , an industry leader ... target cancer stemness pathways, announced that its lead ... Designation from the U.S. Food and Drug Administration ... gastroesophageal junction (GEJ) cancer. Napabucasin is an orally ... stemness pathways by targeting STAT3, and is currently ...
(Date:6/23/2016)... Cancer Foundation (PCF) is pleased to announce 24 new Young Investigator (YI) ... of the Class of 2016 were selected from a pool of 128 applicants ... the Class of 2016 PCF Young Investigators ... ... ...
(Date:6/23/2016)... ... 2016 , ... In a new case report published today in STEM CELLS ... developed lymphedema after being treated for breast cancer benefitted from an injection of stem ... with this debilitating, frequent side effect of cancer treatment. , Lymphedema refers ...
(Date:6/23/2016)... ... June 23, 2016 , ... ClinCapture, ... Pennsylvania Convention Center and will showcase its product’s latest features from June 26 ... presenting a scientific poster on Disrupting Clinical Trials in The Cloud during the ...
Breaking Biology Technology:
(Date:4/28/2016)... and BANGALORE, India , April 28, ... Systems, a product subsidiary of Infosys (NYSE: INFY ... announced a global partnership that will provide end ... use mobile banking and payment services.      (Logo: ... key innovation area for financial services, but it also plays ...
(Date:4/26/2016)... India and LONDON ... Infosys Finacle, part of EdgeVerve Systems, a product ... and Onegini today announced a partnership to integrate ... solutions.      (Logo: http://photos.prnewswire.com/prnh/20151104/283829LOGO ... to provide their customers enhanced security to access ...
(Date:4/19/2016)... 2016 The new GEZE SecuLogic ... web-based "all-in-one" system solution for all door components. It ... the door interface with integration authorization management system, and ... The minimal dimensions of the access control and the ... installations offer considerable freedom of design with regard to ...
Breaking Biology News(10 mins):