RNA isolation / purification Tissue - Human Bone marrow

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.

Start discussion

Found 2 discussions for this experiment

Discussion

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?

Discussion

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?

Share your thoughts or question with experts in your field by adding a discussion!

Found 3 matching solutions for this experiment

Protocol tips
- For RNA to dissolve better and to yield high levels, preheat elution buffer at 95C.

- Addition of β-mercaptoethanol in lysis buffer can be skipped because the phenol will do the job of nuclease inactivation

- To capture more amount of mRNA, modify the amount of lysis buffer :ethanol (from 1:1 to 1:1.5) during the binding step


Downstream tips
- Do vortex for 2-plus minutes during Acid-Phenol:Chloroform extraction step. This will facilitate the removal of tightly bound proteins typically associated with RNA
Upstream tips
- This kit can be used for the isolation of RNA from up to 5x10^5 cells, or up to 5 mg of tissue.
Downstream tips
- Always blank the spectrophotometer with your elution buffer.

- When measuring 260 nm absorbance, a unit of 1 on this scale equals to 40 ng of RNA.
Protocol tips
- To capture more amount of mRNA, modify the amount of lysis buffer :ethanol (from 1:1 to 1:1.5) during the binding step
Can't find the product you've used to perform this experiment? It would be great if you can help us by Adding a product!
Become shareholder Discussions About us Contact Privacy Terms