Does anyone know about which carrier is better for RNA extraction: LPA (linear acrilamide), tRNA or glycogen? I need to analyze samples with low quantities of RNA, make c-DNA and Real time PCR. I heard that tRNA can inhibit c-DNA synthesis... is it true? maybe someone who has worked wtih these can help me! thanks!
I have worked with glycogen and find it to be better than tRNA. Unlike tRNA that shares spectrophotometric properties with other forms of nucleic acid, glycogen does not contribute to the absorption measurements acquired for concentration determinations. Additionally, glycogen can be removed during the 70 % ethanol wash after RNA precipitation thus reducing chances of inhibition in down stream reactions.
Glycoblue is also good, with an added advantage that you can also see the pellets better. I did not notice it making subsequent RT any worse.
The most frequently used RNA carriers are tRNAs (Isolated from bacteria/yeasts). However this is expensive and have some disadvantages. I suggest you to try Ambion Carrier RNA (1 mg⁄mL) Isolation Kits.
There is no need to use carrier RNA. As mentioned before glycogen works perfectly fine and can be removed in the 70% EtOH wash.
As previously mentioned by collegues, according my experience Glycogen works very well and with better performance than tRNA for further experiments that you need to carry out..
For your question: I worked with linear acrilamide and I had good results.
Now, I wish to improve my DNA extraction: Does anyone know about which carrier is better for DNA extraction?
Linear Acrylamide (LPA) works as an excellent carrier molecule in RNA extraction. I never noticed any problems with qRT-PCR in the following steps.
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