RNA isolation / purification Tissue - Mouse Adipose

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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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 4 matching solutions for this experiment

Upstream tips
- This kit purifies RNA from a maximum amount of 30 mg fresh or frozen tissue or 20 mg of tissue that has been stabilized with RNAlater RNA Stabilization Reagent or Allprotect Tissue Reagent can generally be processed.
Protocol tips
- Perform all centrifugation steps at 20–25ºC in astandard microcentrifuge
Downstream tips
- For low A260/A280 value use 10 mM Tris·Cl,* pH 7.5, not RNase-free water, to dilute the sample before measuring purity.
Protocol tips
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- If the RNA yield is low, repeat RNA elution, but incubate the RNeasy Mini spin column on the benchtop for 10 min with RNase-free water before centrifuging.

- All centrifugation steps should be performed at 15–25°C except for phase separation
PureLink™ RNA Mini Kit

Thermo Fisher Scientific

Protocol tips
- Depending on the material you want to isolate RNA from, please refer to the manual for additional reagents/lab equipment that are required.
Downstream tips
- DNase is not supplied with this kit, a DNase kit is available which is compatible with this kit
Protocol tips
- Do not freeze-thaw the DNase more than three times after rehydration.
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