Hi Serra, Since you are new to this field, I would like to mention my way of handling surgical tissues, in detail. Firstly, collect the tissue in medium (Phosphate buffer saline with one protease inhibitor) on ice from the operation theater and transport to the lab immediately. Then, clean the tissues of extra blood mucus etc rapidly using cold fresh medium with protease inhibitor. Divide the tissues into parts, depending on what will be the downstream experiments. As I understand from your question, divide into into 3 parts. For protein studies, I would recommend that you pulverize the tissue (put the tissue in a pre-chilled mortar and pour liquid nitrogen and grind tissue into a fine powder using a pre-chilled pestle). I always prefer to take tissues for RNA extraction almost immediately. If you pulverize carefully, without much shear stress, the pulverized form can be used for all experiments without much damage to the molecules of interest. Storage should always be done at -80 degrees deep freezer. It will also be advisable to always keep a part of the tumor tissue for histology purpose, since later, many a times, you may need to do retrospection.

You are most welcome, Serra

Hi Serra, For RNA purification from freshly collected sample you can use RNAlater: (https://www.thermofisher.com/order/catalog/product/AM7020). In the tissue sample can stabilize RNA for 1 day at 37°C, 1 week at 25°C, 1 month at 4°C, or indefinitely at -20°C.

I have successfully clear intact total RNA from human gut biopsy samples.

For RNA isolation from frozen tissues I really advise the RNAlater-ICE (blue colored liquid). Here you can find furter details: https://www.thermofisher.com/order/catalog/product/AM7030.

I have used it earlier for snapfrozen mice brain tissue RNA isolation, with success. Or for financial purposes, you can you use the more cheaper good old acidic phenol-guanidin-isothyocianate mixture (e.g. Trizol, TRI Reagent) for lysis. Both of the upper lysis solution adding steps further followed by homogenization by mechanical method (stainless steel blades homogenizator or beads). Then use in classical phenol-chloroform-isopropanol purifing method. (Pros: the cheapest, intact total RNA (including rRNAs, miRNAs) can isolated with high yield, Con: toxic organic solvents (need a chemical hood), very problematic to use in high throughput). You can use the Trizol lysate directly in Direct-zol RNA spin column kit (https://www.zymoresearch.com/media/amasty/amfile/attach/_R2060_R2061_R2062_R2063_Direct-zol_RNA_MicroPrep_ver._1.0.0.pdf). I have very good experiences with this kit in human chordoma tissue RNA isolation in the terms of RNA quality and quantity. I have used the Trizole lysate after chloroform phase-separation step in the above kit. To decrease the protein contamination in the further purification steps If you have any questions about RNA isolation from human blood, tissues samples I will answer, if I can. Bests, Tamas