I am using Yeast RNA (Roche 109223) as a carrier for ethanol precipitation of RNA samples. Every time, I have to struggle to dissolve Yeast RNA in DEPC-treated water. Does anybody have a method to easily dissolve and prepare 10 mg/ml Yeast RNA?
I don't know much about yeast tRNA as a carrier for nucleic acids precipitation. However, I have always used glycogen, molecula rbiology grade, which is available from Merck as a 20 mg/ml solution. A single 1-ml tube goes a long way and can be kept indefinitely at -20 °C
Thanks a lot, everybody. I do have glycogen. And, actually, I can obtain sufficient RNAs and DNAs by conventional ethanol precipitation methods, without using carriers. I mainly use Yeast RNA for ethanol precipitation of DIG-labeled RNA probes. And I use Yeast RNA as a component of hybridization buffer for in situ hybridization (Sorry, I forgot to mention this fact in a original question). I know that some manufacturers sell 10 mg/ml Yeast RNA solution, but they are too expensive. I would like to prepare carrier RNA as cheap as possible. Roche 109223 may not be the highest grade, but it has worked as a carrier as well as a blocking agent. However, it is always very difficult and annoying to dissolve RNA in water.
Dissolve 100 mg Worthington Ribonucleic Acid (RNA) in 10 ml 0.1 M sodium acetate buffer, pH 5.0 Stir gently to dissolve to prevent denaturation. Record A260 at 1:50 dilution in 0.10M sodium acetate pH 5.0 to determine mg RNA/ml. mg RNA/ml = A260 x dilution x 0.04. Please see Enzyme manual at http://www.worthington-biochem.com:8080/enzyme-manual/RNA/
for detail.
Hi. I think the following webdite could help you :
http://biscweb.uark.edu/dmcnabb/protocols/acidphenolRNAprep.pdf
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