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Rapid and Cost-Effective RNA Extraction of Rat Pancreatic Tissue
Chapters
Summary September 19th, 2020
The purity and integrity of the isolated RNA is a vital step in RNA dependent assays. Here, we present a practical, rapid, and inexpensive method to extract RNA from a small quantity of undamaged pancreatic tissue.
Transcript
Hi everybody.We have designed, a straightforward, rapid in class descriptive method for efficient RNA extraction from pancreatic tissue.The main advantage of this method is an optimized procedure to extract RNA from minimum amount of tissue due to autolysis.Hi again.Demonstrating of the procedure will be done by Sanaz Dastghaib, that's my PHD student in our lab at Biochemistry Department of Shiraz University of Medical Sciences.First prepare the surgery place, put all requirements under the hood.To inactivate RNase sterilize all the surgery instruments in an oven.Inject ketamine-xylazine intraperitoneally 80/8 milligram per kilogram.Add 1 cc.RNA stabilization reagent into proper microtube to inhabit activation of endonuclease in existed tissues.Immediately place the rat on the surgical board.Head away the surgeon for a midline incision.Then, sterilize all the abdominal surface with 70%alcohol to remove the rat hair.Cut the skin for the surgery.Reshape incision was done to open the abdomen from the pubic area to front legs.Flip the abdominal organs to the left side, to expose the pancreas.Carefully find the pancreas tissue, which is spread in the abdominal cavity.Locate the area under display to find out the pancreas without being confused with the fat tissue.Immediately remove 20 to 30 milligram pancreas from the abdominal cavity in less than two minutes.Put the removed tissue in the micro tube very quickly and cut it with the astride cutter to penetrate the RNA stabilization reagent to separated tissues.Place the microtube in nitrogen tank to freeze immediately.All their microtubes should be kept in the 80
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