I obtained mouse serum samples. My initial plan is to check some proteins using ELISA. However, plans have changed and now I need to check some gene expressions.
My questions are:
1) Can I extract RNA from serum?
2) After the extraction, is there a difference in the regular cDNA synthesis protocol?
3) What is the most efficient kit/protocol you have been using?
Thanks in advance for your time and answers.
https://www.researchgate.net/post/RNA-isolation-protocol-from-Serum-Plasma
Hi Ebru,
First of all we need to know that RNA found in extracellular fluids like serum and plasma is normally fragmented RNA or small RNA , mainly miRNA, and is mostly associated with proteins or contained inside extracellular vesicles. If your target is to purify mRNA from plasma/serum then you have to be aware that this mRNA may not be the full length unlike the original RNA, that you may be getting when isolating RNA from whole blood or tissue samples.
1) Technically yes..!! you can isolate RNA from serum or plasma samples provided you have taken good care of RNAase contamination during sample collection and serum preparation.
2) Post mRNA/total RNA synthesis, there is no difference in cDNA synthesis protocol.
3) There are many kits available in market for isolating RNA from serum samples but I use Qiagen's miRNA easy kit. here is link https://www.qiagen.com/de/products/discovery-and-translational-research/dna-rna-purification/rna-purification/mirna/mirneasy-serum-plasma-advanced-kit/#orderinginformation.
Hope you find it useful.
Your answers as you asked:
1) Yes, it is possible to extract RNA from serum samples. However, it was better to use the buffy coat Layer to obtain a higher RNA concentration. To compensate it, use the high-quality kits to get the highest concentrations available.
2) For cDNA synthesis you can use the comercial kits, which almost all of them have a similar instruction and work well.
3) The kits from outstanding brands (Qiagen, Sigma, Roche, Takara,...) are the best choices. Also, if you need to obtain RNA, protein, and DNA from your samples at the same time, you can use Tri-pure kits.
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