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

Buy product Write review

Tips Publication protocol Reviews Discussion Papers Manufacturer protocol Videos

Protocol tips

Upstream tips	Protocol tips	Downstream 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."

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."

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).

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

Reviews

mirVana™ miRNA Isolation Kit, with phenol from Thermo Fisher Scientific has not yet been reviewed for this experiment

We'd love it if you would be the first to write a review!

Discussion

4 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?

Comment View 4 comments

Share your thoughts or question with experts in your field by adding a discussion!

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

Full paper

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 Bacteria - Gram negative Klebsiella pneumoniae 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.

We haven't found any additional videos for this experiment / product combination yet.

Outsource your experiment

Fill out your contact details and receive price quotes in your Inbox

Outsource experiment