RNA isolated from the priamry neurons

jnilsen jnilsen at pantheon.yale.edu
Fri Mar 2 12:40:56 EST 2001

on 2 Mar 2001 14:42:07 -0000 Jinhongwei (Jinhw at imm.ac.cn) utilized the mandibles to disperse:
#  i am newer comer in this field, i wanna study channels genes from the
# primary cultured hippocampal neuron using RT-PCR.
# if i use Trizol to isolat RNA for neurons,how to deal the cells before
# adding Trizol using scraping cutter or trypsin disgestion

We usually grow the cells in 6-well plates and add1 ml Trizol per well, 
let sit a few minutes and triturate with a pipet before transfering to 
microfuge tubes. Resuspending the RNA pellet in 10 ul of DEPC-water 
usually yields 1.5-2 mg/ml RNA. Alternative you can grow the cells in 
bigger dishes and scrape the cells in PBS, transfer to a centrifuge tube 
and spin down. Then adjust the amount of Trizol based on the size of the 
cell pellet.

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