TRIzol Reagent

RNA isolation / purification Tissue - Mouse Spinal cord

Experiment
RNA isolation / purification Tissue - Mouse Spinal cord
Product
TRIzol Reagent from Thermo Fisher Scientific
Manufacturer
Thermo Fisher Scientific

Protocol tips

Protocol tips
- For low 260/230 readings the best approach is to try washing sample (precipitation) with ethanol to desalt it.

Publication protocol

Total RNA was extracted from each sample individually using TRIzol reagent (Invitrogen, Carlsbad, CA). First-strand cDNA was generated by using a T7-linked-(dT)24 primer. After second-strand synthesis, the double-stranded cDNA was used for in vitro transcription using the ENZO BioArray High-Yield RNA transcript labeling kit (ENZO, Farmingdale, NY). The labeled cRNA was purified and fragmented. Each fragmented, biotin-labeled cRNA sample (20 µg) was hybridized to an Affymetrix mouse 430A chip. After scanning, array images were assessed by eye to confirm scanner alignment and the absence of significant bubbles or scratches. Chips had to meet the following criteria to be included in the data analysis: the number of probe pairs called present had to be greater than or equal to 30%, all spiked-in bacterial controls had to be called present, background had to be <200, the number of outliers had to be <500, and the 3V/5V ratio of GAPDH and actin had to be <2.

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Discussion

Discussion

2 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

3 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 - Mouse Spinal cord using TRIzol Reagent from Thermo Fisher Scientific.

Paper title
Enhanced Functional Recovery in MRL/MpJ Mice after Spinal Cord Dorsal Hemisection
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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for TRIzol Reagent below.

Download PDF Download manufacturer protocol

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