Isolation of total RNA from Kidney biopsies

Peter Hauser (last update 20.11.2002)

Kidney wedge biopsies were obtained right before implantation. A small part (about 12µg) of the wedge biopsy was put into an Eppendorf fube containing 500 µl RNAlater™ (Ambion, Austin, TX). Samples where stored at +4°C over night and kept at -80°C for long-time storage.


  • Put biopsy into 1 ml TRIzol®, grind tissue till a homogeneous mass emerges
  • If frozen tissue is used; thaw on ice and remove RNAlater® with pipette
  • Transfer grinded biopsy to an Eppendorf tube and add 200 µl chloroform
  • Incubate in orbital shaker for 3 minutes
  • Centrifuge (20 minutes, 10000 g, 4°C)
  • Transfer the aqueous (upper) phase to a new Eppendorf tube
  • Precipitate RNA by adding an equal amount (~550 µl) of isopropanol
  • Incubate 10 minutes at room temperature
  • Centrifuge precipitation mix (20 minutes, 10000 g, 4°C)
  • Wash pellet in 1ml 70% ethanol
  • Centrifuge again (10 minutes, 10000 g, 4°C)
  • Air dry pellet at room temperature for 15 minutes
  • Resuspend in 10µl nuclease free H2O


7 ml Tenbroeck tissue homogenizer Wheaton Science Products, Millville, NJ
TRIzol Reagent® GibcoBRL Gaithersburg, MD #15596-026
Chloroform Merck # 1.59129
Isopropanol Sigma # 405-7
Ethanol Merck # 1.00983.1011
Nuclease free Water GibcoBRL Gaithersburg, MD #10977-015