mirVana™ miRNA Isolation Kit, with phenol

RNA isolation / purification Tissue - Human Esophagus

Experiment
RNA isolation / purification Tissue - Human Esophagus
Product
mirVana™ miRNA Isolation Kit, with phenol from Thermo Fisher Scientific
Manufacturer
Thermo Fisher Scientific

Protocol tips

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.

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Discussion

Discussion

4 years ago

Author: Paul G. Macon United States

RNA isolation from tissue

How do I extract RNA from animal tissue without using liquid nitrogen? I tried the RNA extraction by using the TRIzol reagent and I homogenize the tissue using polytron homogenizer at room temperature for 30secs is this correct?

Discussion

5 years ago

Author: Aaron Stege Netherlands

Problem in phase separation after using serum/plasma kit

I used a serum/plasma kit for my serum samples. After the phase separation the samples should have 3 phases: a colourless aqueous phase, a white interphase and a red organic phase. However, in some of my samples there was no aqueous phase unless I wait for an extended period of time. How can I circumvent this problem?

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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.

Paper title
Single cell RNA-seq reveals profound transcriptional similarity between Barrett’s oesophagus and oesophageal submucosal glands
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Manufacturer protocol

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

Download PDF Download manufacturer protocol

Videos

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