[northern] Detact of tRNA aminoacylation and deacylation in p38KO MEF cells containing 35-S Methionine.

[Seung Jae Jeong]

Detact of tRNA aminoacylation and deacylation

in p38KO MEF cells containing 35-S Methionine.

2001 Joong-Won Lee

 

 

 

 

1.prepare methionine-free DMEM containing 10% dialyzed FBS.

 

2.washing the dish with 1X PBS two times and change the medium by prepared in step 1(Methionine-free medium) 4-5ml each 100Ø dish.

 

3.incubation for 30min at 37C⁰

 

4.add [35-S] Met (50-100uCi)

 

5.incubation for 30min at 37C⁰

 

6.washing with 1X PBS and harvest

 

7.total RNA prep.(two sample : dissolved in D.W. or 10mM NaOAc)

 

8.deacylation tRNA and northern blots

 

 

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