Extraction method for acute pancreatitis model animal pancreas tissue RNA

A technique for acute pancreatitis and model animals, which is applied in the field of molecular biology and can solve the problems of poor RNA extraction from pancreas tissue of animals with acute pancreatitis

Inactive Publication Date: 2007-10-24
XI AN JIAOTONG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The above method is suitable for the extraction of RNA from pancreatic tissue of normal animals, but the extraction effect of RNA from pancreatic tissue of animals with acute pancreatitis is poor

Method used

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  • Extraction method for acute pancreatitis model animal pancreas tissue RNA
  • Extraction method for acute pancreatitis model animal pancreas tissue RNA
  • Extraction method for acute pancreatitis model animal pancreas tissue RNA

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0014] 1. Preparation of experimental drugs and disposal of experimental supplies

[0015] 1) Preparation of experimental drugs

[0016] Lavage fluid: HEPES 25mM, collagenase IV 40U / ml, soybean trypsin inhibitor 0.1mg / ml, NaCL 100mM, KCL 5mM, MgCL 2 1mM, CaCL 2 1mM, D-dextrose 14mM, glutamine 2mM, 2% (w / v) BSA, prepared with sterile saline, sterilized on the filter membrane in 95% O 2 , 5% CO 2 , Incubate at 37°C for 2 hours for later use.

[0017] Soybean trypsin inhibitor (0.2mg / ml): Prepared with sterile saline and stored at 4°C.

[0018] 75% ethanol: Prepared with RNase-free deionized water treated with DEPC and stored at 4°C.

[0019] TE solution: Use DEPC-treated deionized water without RNase to dilute the sterile packaged TE concentrate.

[0020] 2) Handling of experimental supplies

[0021] Deionized water was added to DEPC with a concentration of 0.1%, heated at 37°C for 12 hours, and subjected to high temperature treatment at 125°C for 2 times for 30 minutes each time to...

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Abstract

The invention discloses a total RNA extracting method of pancreatic tissue of acute pancreatic disease animal, which comprises the following steps: preparing lavage liquid; breeding film after degerming for 2h in the 95% O2 and 5% CO2; narcotizing the ill animal; plugging pipe into main pancreatic duct from the opening of duodenum; injecting fitful quantity of lavage liquid; cutting the pancreatic tissue; pruning in the soy parenzyme inhibitor under 4 deg. c; washing; placing in the TRlzol cracking liquid to homogenize at 4 deg. c; extracting the total RNA of pancreatic tissue according to the instruction of TRlzol cracking liquid agent box; detecting the density and the rate of OD260 and OD280 through spectrophotometric method; estimating the density; identifying the completeness integrity through 1% agarose gel electrophoresis.

Description

Technical field [0001] The present invention relates to the fields of molecular biology such as gene cloning, gene function and transgene of the pancreas, and more specifically to a method for extracting pancreatic tissue RNA from acute pancreatitis model animals. The method is also applicable to total pancreatic tissue RNA from other acute pancreatitis model animals Extraction. Background technique [0002] Acute pancreatitis is a common and severe clinical disease with many complications and difficult treatment. At present, the investigation of its pathogenesis has reached the level of gene transcription. The extraction of total RNA from pancreatic tissue of acute pancreatitis model animals is a basic technique for experiments such as cDNA library construction, Northern hybridization, and RNA difference display. Compared with normal animal pancreatic tissue, it is more difficult to extract total RNA from pancreatic tissue of acute pancreatitis model anima...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07H1/06C07H21/02
Inventor 李宗芳夏先明张澍杨军黄辰王波张爱军
Owner XI AN JIAOTONG UNIV
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