TRIzol Reagent
RNA isolation / purification Tissue - Human Thyroid gland
Protocol tips
| Protocol tips |
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| - For low 260/230 readings the best approach is to try washing sample (precipitation) with ethanol to desalt it. |
Publication protocol
With the exception of 17 samples which were excluded because of poor RNA quality, total RNA was extracted from frozen thyroid tissues with TRIzol reagent (Invitrogen, Carlsbad, CA), the concentration determined on a Nanodrop spectrophotometer (Nano-Drop, Wilmington, DE), and quality assessed with the Agilent RNA 6000 Nano Kit on an Agilent 2100 Bioanalyzer (Agilent Technologies, Milan, Italy). For each sample, 300 ng of total RNA was reverse transcribed to synthesize cDNA and biotinylated cRNA according to the Illumina TotalPrep RNA amplification protocol (Ambion; category n. IL1791). Hybridization of 750 ng of cRNA on Illumina HumanHT12 v4.0 Expression BeadChip array (Illumina Inc.), staining and scanning were performed according to the standard protocol (Illumina Inc.). BeadChip was dried and scanned with an Illumina HiScanSQ system (Illumina Inc.).Full paper Login or join for free to view the full paper.
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Discussion
Discussion
4 years ago
Author:
Paul G. Macon
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
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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Manufacturer protocol
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