Heh. This is one of those questions that just raises far, far more questions.
Fundamentally human tissue isn't significantly different from mouse or rat or horse or pig or any mammalian tissue, about the only thing you can usually rely on is that you'll have less of it. Unless you're using tissue from surgery, or from the recently deceased.
Generally speaking you can't go too far wrong with just "Trizol", but preparation of the material BEFORE you get to that point can vary quite a bit.
So. Questions that will help us answer:
Which tissues?
How much tissue?
How fresh is the tissue?
What condition is the tissue in?
Are you hoping to use the same tissue sample for other investigations?
RNAqueous®-Micro Kit is a convenient low elution silica column. It is very Fast and you can consistently isolate RNA from even a micro-sized samples. This method only take 15 minutes only. Moreover, you can also use TRIzol® Plus RNA Purification System.
Dear Haritha Doddam Reddy
As John Hildyard mentioned, we need to know some information about your tissue and .... but I share protocols that I usually use them for extracting RNA from breast , thyroid& colorectal tissues (RNX plus kit & TRIZOL kit)
TRIZOL kit procedure:
1. At first, freeze tissue samples immediately upon collection.
2. Add 1mL TRIZOL kit per 50-100 mg of tissue.
3. Homogenize tissue using a glass-Telfon for 5 min.
4. Incubate the homogenized sample for 5min at room temperature (20-25 oC)
5. Add 200 microliter of chloroform per 1mL of TRIZOL.
6. Shake tube vigorously by hand for 20 sec.
7. Incubate for 5 min at room temperature.
8. After incubation, centrifuge the sample at 12,000 xg for 15min at 4 oC.
9. Remove gently the aqueous phase and place it into a new tube.
10. Add 500 microliter of 100% isopropanol (4-7 oC) to the aqueous phase .
11. Incubate it at room temperature for 15min.
12. Centrifuge at 12,000xg for 15min at 4 oC.
13. Remove and discard supernatant from the tube and wash the pellet with 1mL of 75% ethanol.
14. Centrifuge the tube at 7,500xg for 8 min at 4 oC.
15. Remove and discard supernatant from the tube and dry the RNA pellet for 10 min at room temperature.
16. Resuspend RNA pellet in RNase-free water or DEPC water by passing the solution up and down several times through a pipette tip (amount of water(10-50microlitter) is dependent on pellet size).
17. Store RNA solution at -80 oC.
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