mirVana™ miRNA Isolation Kit, with phenol

RNA isolation / purification Tissue - Human Bone marrow

Experiment
RNA isolation / purification Tissue - Human Bone marrow
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 yield 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

Total RNA was extracted using mirVana Isolation Kit (Applied Biosystem, Foster City, CA). We obtained good-quality RNA from all the samples, with a mean yield of 16.2 ± 11.3 μg of total RNA. Reverse-transcription was performed using 5 µg total RNA with SuperScriptIII (Roche, Basel, Switzerland). Real-time quantitative RT-PCR was performed to study the expression of selected genes with TaqMan Gene Expression Assays (Applied Biosystems). Results were analyzed with RT2 Profiler PCR Array software (Qiagen).

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Reviews

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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 Bone marrow using mirVana™ miRNA Isolation Kit, with phenol from Thermo Fisher Scientific.

Paper title
Autologous Pancreatic Islet Transplantation in Human Bone Marrow
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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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