mirVana™ miRNA Isolation Kit, with phenol

RNA isolation / purification Bacteria - Gram negative Klebsiella pneumoniae

Experiment
RNA isolation / purification Bacteria - Gram negative Klebsiella pneumoniae
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 a higher 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. This will facilitate the removal of tightly bound proteins typically associated with RNA."

Publication protocol

Samples were grown in CA-MH broth to an OD600 of 0.6 before addition of RNAprotect bacteria reagent and RNA extraction using the mirVana RNA isolation kit (Ambion). A total of 500 ng of RNA was converted to cDNA using Superscript II (Life Technologies) followed by qRT-PCR using SsoFast Sybr green (Bio-Rad).

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Reviews

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Discussion

Discussion

5 years ago

Author: Paul G. Macon United States

Some help with RNA isolation using Trizol

Hello! I used Trizol to extract total RNA from in-vitro cultured bacteria (1 X 10^8 cells). After phase separation, I mixed ~0.4 ml of the upper phase which contains RNA with 0.5 mL cold isopropanol. However, the amount of RNA when measured in Nanodrop was very low. In addition, the ratio between 260 and 230 was around 0.1 to 0.5. Is there a chance that my sample was contaminated by the Trizol reagent? When I collected the aqueous phase I made sure to not touch the lower phase. What should I do?

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Papers

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

Paper title
IS Element Integration, a Novel Mechanism for Rapid Emergence of Tigecycline Nonsusceptibility in
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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

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