mirVana™ miRNA Isolation Kit, with phenol

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- For RNA to dissolve better and to yeild high levels, preheat elution buffer at 95C. - Addition of β-mercaptoethanol in lysis buffer can be skipped because the phenol will do the job of nuclease inactivation - To capture more amount of mRNA, modify the amount of lysis buffer :ethanol (from 1:1 to 1:1.5) during the binding step	- Do vortex for 2-plus minutes during Acid-Phenol:Chloroform extraction step. This will facilitate the removal of tightly bound proteins typically associated with RNA.

Protocol tips
- For RNA to dissolve better and to yeild high levels, preheat elution buffer at 95C. - Addition of β-mercaptoethanol in lysis buffer can be skipped because the phenol will do the job of nuclease inactivation - To capture more amount of mRNA, modify the amount of lysis buffer :ethanol (from 1:1 to 1:1.5) during the binding step
Downstream tips
- Do vortex for 2-plus minutes during Acid-Phenol:Chloroform extraction step. This will facilitate the removal of tightly bound proteins typically associated with RNA.

Publication protocol

Tissue fragments were processed using the mirVana™ miRNA Isolation Kit (ThermoFisher) according to manufacturer’s guidance. Total RNA was enriched using ribodepletion (Ribo-Zero, Illumina®) prior to cDNA conversion. Second strand DNA synthesis incorporated dUTP. cDNA was end-repaired, A-tailed and adaptor-ligated. Samples then underwent uridine digestion. The prepared libraries were size-selected and multiplexed before 75 bp paired end sequencing using the Illumina® HiSeq 4000 platform.

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing RNA isolation / purification Tissue - Human Esophagus using mirVana™ miRNA Isolation Kit, with phenol from Thermo Fisher Scientific.

Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for mirVana™ miRNA Isolation Kit, with phenol below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing RNA isolation / purification Tissue - Human Esophagus using mirVana™ miRNA Isolation Kit, with phenol from Thermo Fisher Scientific. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.