$120/kit of 50 preps plant total RNA Isolation Kit with patent O-ring column. Pure, wash buffer free plant RNA, better A260, 280, 230. Free shipping on $200
DNALand Scientific Plant total RNA Isolation Kit with patented O Ring Column all researchers to isolate total RNAs from 1 g plant material or 1 x 108 cells.
* Northern blotting
* In vitro translation
* cDNA synthesis
1. Grind 1 g (or less) plant sample under liquid nitrogen to a fine powder and transfer to a new tube.
2. Add 4.5 ml of RX Buffer or PRX Buffer (-ME added) to the tissue powder and vortex vigorously. In most cases RX Buffer is the buffer of choice to lyse plant tissue. However, plant tissues contain sticky secondary metabolites (for example, maize with milky endosperm or mycelia of filamentous fungi), PRX Buffer is used instead.
3. Apply lysate to the Shearing tube sitting in a Collection tube and centrifuge at full speed (3,000 rpm or 2,500 x g) for 2~10 minutes. Transfer flow-through sample from the Collection tube to a new tube.
Avoid pipetting any debris and pellet in the collection tube. To centrifuge for 10 minutes will enhance the extracted RNA quality much.
4. Add 2.3 ml (about half of the sample volume) 98-100% ethanol to the clear lysate and mix by pipetting.
If sample lysate is lost during the preparation, reduce ethanol volume proportionally.
5. Apply 6.8 ml of the ethanol added sample (including any precipitate) from step 4 to a Plant Total RNA Maxi Column sitting in a Collection Tube, close the cap, centrifuge at full speed for 3 minutes, and discard the filtrate.
If the solution remains above the membrane, centrifuge again for another 5 minutes.
6. Repeat step 5 for rest of the sample.
7. Wash the column once with 5 ml of WF Buffer by centrifuging at full speed for 3 minutes and discard the filtrate.
8. Wash the column twice with 5 ml of WS Buffer by centrifuging at full speed for 3 minutes and discard the filtrate.
Add 100 ml of ethanol (98-100%) to the WS Buffer bottle when first open the bottle.
9. Centrifuge at full speed for 3 minutes to remove traces of WS Buffer.
Residual ethanol may inhibit reverse transcriptase activity.
10. Transfer the column to a RNase-free 15 ml Elution Tube (provided), add 500 l of RNase-free ddH2O, Stand the column for 5 minutes. Centrifuge for 1-2 minutes to elute DNA.and centrifuge at full speed for 5 minutes to elute RNA.
Store RNA at -70C.
The shipping cost for each case ordered is $9.50, but will be automatically waived at checkout on orders of $200 and more.