We have crushed plant samples in micro-tube (2 ml) which previously filled in until less than half. After adding TRI reagent, chloroform, and centrifuging, we do not get aqueous phase!, only orange one containing protein!
What is the best solution?
Thanks
Hello Seyed,
Yes the volume/amount of sample is very important for good RNA prep. For Trizol/Tri-reagent protocols, the maximum recommended amount of tissue sample is 100mg/ml of the reagent. For tissue homogenization, either use a sterile mortar and pestle with liquid nitrogen or homogenize the sample in trizol/tri-reagent. Typical quantity of RNA obtained following trizol method is around 2-3 micrograms/microliter. Use DEPC treated plasticware, and the recommended precautions to avoid RNAse contamination.
Hi Dheeraj,
thank you for your answer.
I have already crushed samples in -80 which are more than 100 mg in each tube. The question is whether I should transfer 100 mg of each collected tube into new tube or ? If so, maybe I will lose my RNA!
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Biomolecules
- Nucleic Acids
- RNA