RNA isolation / purification Tissue - Human Adrenal glands

Isolating RNA from tissues and paraffin embeded tissue samples can be challenging due to cross-linking of biomolecules and fragmented nucleic acids. The best solution is to slice the tissues into smaller pieces and make a homogenate solution (using tissue homogenizer or grinding liquid nitrogen frozen samples) in presence of RNAse inhibitors. The homogenization process should be carried out on dry ice to maintain the intigrity of RNA

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1 year ago

1 year ago by 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?

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1 year ago

1 year ago by 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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Found 3 matching solutions for this experiment

Upstream tips
For lysis/cell disruption check the protocol beforehand to see which reagents/lab materials are needed
Downstream tips
Minimal DNA is left over using this kit, however if DNA is undesired an additional RNase free DNase treatment is warranted.
Downstream tips
Ensure EtOH is completely evaporated off of the column prior to elution. Adjust time from 1min to 5 min at 60C.

Use water to elute the RNA that is warmed to ~60C.
TRIzol Reagent

Thermo Fisher Scientific

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