Tuesday September 25, 2007
The Ribo-Sorb Nucleic Acid Extraction Kit is intended for the isolation and purification of RNA/DNA from plasma, serum, whole blood, liquor, amniotic liquid, tissue, urine, feces, bronco aspirates and other biological materials. This kit can be used to simultaneously isolate RNA and DNA. I have used it mostly for RNA extraction from patient serum or plasma. However, the kit manufacturer recommends extraction from plasma over serum because all publications show that plasma retains more virus than serum.
I have found Ribo-Sorb nucleic acid extraction to be easy to perform and it has generated high yields of RNA. The procedure is very easy; even those who have no experience with RNA extraction can use it. Before starting an extraction, you need to bring all the reagents at room temperature, and if you see any crystals in the lysis or washing buffer, you must warm it to 60°C until crystal disappear (usually10 minutes). As always, it is a good idea to include positive and negative controls, if you have them, in order to monitor the kit’s quality as well as the cleanliness (i.e. no RNases) of the work area. The procedure takes just 40 minutes. First, label a 1.5 mL polypropylene tube and add 450 uL of lysis buffer plus 100 uL of your sample (serum, plasma, whole blood, etc). Vortex and briefly spin the sample, then add 25 uL Sorbent (a 30% Silica dioxide suspension in deionized water, included in the kit). Incubate at room temperature for 10 minutes, vortexing periodically. Centrifuge the sample at 10000g for one minute, discard the supernatant, wash the pellet with 400 uL washing solution, vortex, centrifuge at 10000g for one minute. Again discard the supernatant and add 500 uL ethanol (70%). Repeat ethanol wash once more then wash the pellet with 400 uL acetone. Vortex and centrifuge for one minute at 10000g. Using a micropipette or vacuum aspirator with a plugged aerosol barrier tip, carefully remove and discard the supernatant from each tube without disturbing the pellet. Incubate the tubes with the cap open for 10 min at 56°C. Add 50 uL of RNA eluent and incubate at 56°C for 10 minutes. Finally, centrifuge at 10000g. The supernatant contains the RNA, which can be used in experiments or stored at -80°C
We have had good success with this kit. The instructions are very easy to follow and all the necessary reagents are supplied, except ethanol and acetone. The only equipment required is a small tabletop centrifuge. This kit is cheaper than the other kits in the market, like those from Qiagen, Invitrogen, and Gentra. Overall, this kit is less expensive, less laborious, does not require additional consumables, allows extraction from various types of biological samples, saves time, and results in a high quantity and quality of RNA.
Note: This kit is only for research use.
Imtiaz Alam
Manager
Molecular Diagnostics
Global Biotech Pvt Ltd
Pakistan