Frozen Tissue Processing for Proteomics and Genomics
Scientist
on
July 3, 2024
This blog post was written by Cureline, a global translational and precision medicine CRO providing HBS biobanking and laboratory services for accelerated drug R&D and Dx development since 2003. Their services are available on the Scientist.com marketplace.
Fresh-Frozen Tissue Cryopulverization Service
Frozen tissues can be difficult to handle resulting in degradation of valuable biomarkers. Cureline Molecular Services provides the tissue cryopulverization service that converts a regular frozen tissue sample into multiple user-friendly aliquots with a defined cryopowder weight.
The cryopulverized sample format is highly recommended for sensitive downstream analytical methods. Preparation of frozen tissue powder aliquots allows:
- retaining biomolecule integrity found in original tissue sample
- improving tissue solubility in lysis buffers
- increasing yield and integrity of nucleic acids and proteins
- better extraction efficiency of biomarkers for multiple downstream analyses (RNA-Seq, DNA-Seq, PCR, Proteomics, Transcriptomics, etc.)
- generating multiple aliquots from one frozen tissue sample for a variety of downstream applications
We can use in-house or customer-provided tissue specimens for processing. Frozen tissue is prepared using a proprietary NO-THAW (TM) tissue cryopulverization protocol. Tissue powder is weighed in frozen state and prepared as 100 mg standard aliquots (other size is available upon request). Aliquots are shipped on dry ice and ready for downstream applications!
For example, if you have a 0.5 gram tissue chunk, we can prepare up to 5 aliquots with 100 mg of powder. Tissue aliquots are ready for DNA, RNA and protein extraction using common protocols and commercially available kits. Just add an appropriate lysis buffer and proceed with the extraction protocol.
RNA Extraction from Fresh-Frozen Tissue
RNA degradation during extraction from frozen tissue samples can be a significant challenge. When tissues are frozen and subsequently thawed, the mechanisms that protect RNA from degradation by RNases in living cells can be compromised. This can lead to rapid RNA degradation.
Cureline Molecular Services developed a proprietary protocol for RNA isolation from fresh-frozen tissues without the loss of RNA integrity.
The unique RNA isolation protocol allows:
- retaining RNA integrity (RIN) found in original tissue samples
- better extraction efficiency and yield from any tissue type
- smaller tissue weight to isolate sufficient quantity of RNA
- suitability for sensitive downstream applications (RNA-Seq, etc.)
Cureline, Inc. is a global precision and translational medicine CRO providing preclinical and laboratory services to the biopharmaceutical industry since 2003. Cureline Group offers a full range of ISO-certified, CAP-compliant clinical research services (Biobanking, Histo/Digital and molecular pathology, small animal and POC validation) for biopharma researchers conducting preclinical and biomarker/companion diagnostics development projects. Cureline headquarters, based in California (USA), oversees global clinical operations and regulatory affairs in 20+ countries (US, EU, Eastern Europe, China, India, and South America). A preclinical studies center is in Vilnius, Lithuania.