TRIzol Reagent
RNA isolation / purification Tissue - Human Hippocampus
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
To extract total RNA, 1 ml of TRIzol Reagent (Life Technologies, Foster City, CA, USA) was added per 75 mg of frozen tissue samples, homogenized and then further processed according to the manufacturer’s instructions. The RNA integrity number (RIN) mean in both the control and patient groups was similar: 6.68 ± 0.9441 (n = 5) and 6.155 ± 0.2484 (n = 11), respectively. Due to the fact that the RNA was unavailable, the RIN was not evaluated for three patient samples: TLE 03, TLE 11 and TLE 13. Subsequently, 1 μg of total RNA of each sample was reverse transcribed into cDNA using 200 U of Superscript III Reverse Transcriptase (Life Technologies) and 3 μg of Random Primers (Life Technologies) according to the manufacturer’s instructions.Full paper Login or join for free to view the full paper.
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Discussion
Discussion
5 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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