RNA isolation / purification Tissue - Human Placenta

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.

6 Matching solutions
2 Discussions

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Discussion

4 years ago

4 years 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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Discussion

5 years ago

5 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 6 matching solutions for this experiment

RNeasy Mini Kit

Qiagen

Upstream tips	Protocol tips	Downstream tips
	- To yield high RNA, give the column an extra wash with both RW1 and RPE buffers (3 total washes with each buffer).	- 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.

Protocol tips
- To yield high RNA, give the column an extra wash with both RW1 and RPE buffers (3 total washes with each buffer).
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.

Manufacturer protocol 1 Relevant paper

RNAprep pure Kit

Tiangen

Upstream tips	Protocol tips	Downstream tips
- This kit is designed for tissue samples of 20 mg	- β-Mercaptoethanol (β-ME) must be added to Buffer RL before use. The final concentration of β-ME is 1%

Upstream tips
- This kit is designed for tissue samples of 20 mg
Protocol tips
- β-Mercaptoethanol (β-ME) must be added to Buffer RL before use. The final concentration of β-ME is 1%

Manufacturer protocol Publication protocol 1 Relevant paper

NucleoSpin® RNA

Macherey Nagel

Upstream tips	Protocol tips	Downstream tips
- Aliquot rDNase and store at -20 °C.	- Dry the columns very well after the ethanol wash by adding an additional 2' >10000 rpm centrifuge with no buffer.

Upstream tips
- Aliquot rDNase and store at -20 °C.
Protocol tips
- Dry the columns very well after the ethanol wash by adding an additional 2' >10000 rpm centrifuge with no buffer.

Publication protocol 1 Relevant paper

TRIzol Reagent

Thermo Fisher Scientific

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

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

Manufacturer protocol 1 Relevant paper

MagNA Pure Compact RNA Isolation Kit

Roche Lifesciences

Upstream tips	Protocol tips	Downstream tips
- This kit can be used for the isolation of RNA from up to 1x10^6 cells

Upstream tips
- This kit can be used for the isolation of RNA from up to 1x10^6 cells

Manufacturer protocol Publication protocol 1 Relevant paper

TRI Reagent® MRC

Molecular Research Center, Inc.

Upstream tips	Protocol tips	Downstream tips
This kit purifies RNA from up to 50-100 mg tissue

Upstream tips
This kit purifies RNA from up to 50-100 mg tissue

Manufacturer protocol Publication protocol 1 Relevant paper

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