User:Vijay/Protocol/microarray-rna

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Materials

  • RNA Protect (Bacteria) - Qiagen
  • Trizol - Invitrogen
  • Fast Blue - MD (Qbiogene)
  • Chloroform
  • DEPC water
  • 75% ethanol

Procedure

  • Cell Harvest
  1. Collect 2ml culture and add 4mml of RNA protect (Qiagen) in 15 ml Falcon tube.
  2. Vortex for 5sec, incubate room temperature for 5 min.
  3. Centrifuge for 10min at 7300rpm.
  4. Dab on paper towel (up side down).
  5. Freeze.
  • RNA Isolation
  1. Resuspend the pellet in 1ml Trizol (pipette up and down to resuspend) and transfer to FastB tube.


Procedure

  1. SDS-PAGE and TRANSFER done as per our lab protocol

Buffers

1M KH2P04

(No autoclave, no filter, just make it and use it.... in 100ml glass bottle)
13.6gms in 100ml DEPC water

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