I am trying to get good quality RNA isolated from various brain tissues of monkey. can I get some expert views for RNA isolation for this RNA-Seq? What kit or method is best suitable for RNA isolation from animal tissues for this.
Thanks
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https://www.thermofisher.com/ru/en/home/life-science/dna-rna-purification-analysis/rna-extraction/rna-sample-extraction/tissue-rna-extraction.html
Any traditional column-based kit would be fine. Would recommend a column for extracting both microRNA and mRNA as you can just isolate "total RNA", use the mRNA for RNAseq and the remaining RNA can be used for later studies (i.e. microRNA). You will also have the option of doing the optional on-column DNase digest although i have personally never had issue with DNA contamination in RNA but from what i hear it does happen which can give headachesfor RNA-seq experiments.
https://www.qiagen.com/us/shop/sample-technologies/rna/mirna/mirneasy-mini-kit/#orderinginformation
Search the net and you will find thousands of references!!
just one here
https://www.thermofisher.com/ru/en/home/life-science/dna-rna-purification-analysis/rna-extraction/rna-sample-extraction/tissue-rna-extraction.html
A commonly used kit is the RNeasy kit from Qiagen. It generates high-quality total RNA, which can be used directly for RNA-seq. For animal tissues, the key step in the process is homogenization. Make sure that your tissue is well homogenized for good yield. Pay attention to all the potential aspects of the process that may lead to degradation, which are listed in the kit's user manual. Compared to DNA, RNA is much easier to be degraded by RNA nuclease which seems to be omnipresent.
To inhibit the degradation, put some beta mercapto ethanol (BME) to lysis buffer (1~2% volume of lysis buffer). BME will deactivate enzyme which can cause RNA degradation.
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