RNA isolation / purification Tissue - Human Skin

Isolating RNA from tissues and paraffin-embedded 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 integrity 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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2 years ago

2 years 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

NucleoSpin® RNA XS

Macherey Nagel

Upstream tips
- This kit is for the isolation of RNA from very small samples (up to 5 mg)
Protocol tips
- For adsorption measurement, use 5 mM Tris pH 8.5 as diluent
Downstream tips
- Do not let the flow-through touch the column outlet after the second Buffer RA3 wash. Be sure to centrifuge at the corresponding speed for the respective time in order to remove ethanolic Buffer RA3 completely.
Downstream tips
- Include DNAse treatment for 15-20min.

- Ensure EtOH is completely evaporated off of the column prior to elution. Adjust time from 1min to 5 min at 60`C.

- Use water to elute the RNA that is warmed to ~60`C.
PicoPure™ RNA Isolation Kit

Thermo Fisher Scientific

Upstream tips
- Can be used when there is a very less amount of starting material
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