I am looking for a kit that can yield good quality RNA from small sample size (around 10 000 - 15 000 cells). I have tried with Trizol, but so far have been unsuccessful in getting good enough RNA for RNA seq.
Product Name: RNeasy Micro Kit Micro RNA Extraction Kit
Brand and Number: Qiagen 74004
I would also propose an RNa isoloation kit, maybe the RNeasy Micro Kit or other kits of comparable quality but lower price, e.g. try roboclone products!
Hello,
I do think that your RNA prep issue remains not in the purification kit but in the small number of cells you use. Is RNA ses the only approach you want to test? If qPCR should be an option, I would recommend to use life technologies Cell to CT kits..... If RNA ses mandatory and if price matters, I would recommend Macherey Nagel nucleospin kits rather than Qiagen ones. Approximatively same yields but much more cheaper. One of the advantage of nucleospin kit is that you can, if you need but I do not recommend it, enrich your sample prep for small ncRNA.
Hope this will help
Hi Pengfei and Andreas, thanks for the recommendation. Have you used the kits to extract RNA from small sample size successfully for RNA seq? I have noticed that some researchers are concerned that carrier RNA in these kits can interfere with RNA seq. I'm unsure how carefully I should look into it.
Hi Arnaud, yes I need to do RNA seq for my samples. So far I have been doing qPCR using Trizol extraction, but the quality seems to be not good enough for RNA seq. Have you been able to obtain RNAseq quality yields with the nucleospin kits for small volume? Thanks for your recommendation.
Yes we also performed RNA seq but with somewhat higher amounts of cells...however, with good results
Hello Ayon, I do not use cells as a matrix for my molecular biology assays. In a previous company we were use to use nucleospin kits to run either qPCR and RNA seq experiments from low amounts of tissues fresh frozen or FFPE slides. We got quite good yield of good quality RNA (RIN >7).
for your second question, you can use glycogen as RNA carrier. it will not interfere with RNA Seq.
Best
Arnaud
You could also try Zymo's Quick-rna kits. They're cheaper than Qiagen and have higher yields. https://www.zymoresearch.com/collections/quick-rna-kits
Here is some literature in case you are curious:
https://files.zymoresearch.com/literature/pdf/b1003_rna_brochure.pdf
Hope this is helpful and good luck.
Hi Jack,
Thanks for the suggestion, have you personally used this kit for RNAseq?
Better trying RNAzol from sigma. Find the attachment for further details.
Ayon Hassan
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