nt pellet to a fresh 50 ml tube without any carryover of the cell debris. If the pellet is loose, the supernatant should be filtered through an RNase-free Miracloth (Callbiochem).

Add 18 ml 70% ethanol to lysate and completely mix via very gentle, repeated inversion.

Add 4 ml well-mixed Perfect RNA Binding Matrix Solution and mix via gentle inversion.

Place two Perfect RNA Maxi Spin Columns into 50 ml centrifuge tubes and transfer lysate/Binding Matrix mixture to two Spin Column assemblies.
Note: A modified preparation procedure for difficult tissues is described in the detailed protocol Step 6.

Centrifuge for 5 to 10 min. at 2,000 x g. Discard filtrate.

Add 10 ml Wash Solution I to each Spin Column and centrifuge for 5 min. at 2,000 x g. Carefully transfer Spin Columns to fresh centrifuge tubes without splashing Wash Solution I.

Add 10 ml Diluted Wash Solution II (ethanol not provided) to each Spin Column and centrifuge 5 min. at 2,000 x g. Carefully transfer dry Spin Columns to RNase-free 50 ml Collection Tubes (provided).

Add 1 ml Molecular Biology Grade Water to each Spin Column and vortex assemblies briefly.

Incubate at 50C for 5 min.

After incubation, vortex for 5 sec. and immediately centrifuge for 5 min. at 2,000 x g. Store recovered total RNA at 80C.

Results

About 200 g RNA of each were obtained from 1 g source material of Arabidopsis leaves, flowers, or pods, while 5 g yielded about 800 g RNA, and 10 g yielded 850 g RNA. With respect to maize, the yields were somewhat lower: 180 g, 560 g, a
'"/>

Source:

Page: All 1 2 3 4 5 6
Related biology technology :

1. Simple, Sensitive, and Rapid Detection of FLAG -Tagged Fusion Proteins
2. A Simple and Rapid Technique to Purify High-Quality Midiprep DNA
3. Simple Purification of DNA from Plasmid Minipreps, PCR Amplifications and Agarose Gels
4. Simple, Sensitive Isotyping of Mouse Monoclonal Antibodies
5. Simple Isolation of RNA from Tissue and Cultured Cells
6. HSVision Molecular Beacon Detection Module Rapidly Detects Herpes Simplex Virus DNA
7. The Fastest, Simplest, and Most Effective Way to Remove DNA Contamination
8. Simple Approaches for Optimization of RT-PCR
9. Detection of Morphine and Morphine-Glucuronide in Saliva or Urine with a Rapid and Simple Method Without Derivatization on the Varian 500-MS
10. Gram Quantities of MAb Produced with Simple Bioreactor in Serum-Free Perfusion Culture Replacing Ascitic Fluid Production
11. CastAway Precast Gels for Rapid Automated DNA Sequence Analysis