Difference between revisions of "4EBP1/2 Genotyping"

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==PRIMERS==
 
==PRIMERS==
 
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'''4EBP1'''
"'4EBP1"'
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*4EBP1- Fwd: GAT GGA GTG TCG GAA CTC ACC (1X)
 
*4EBP1- Fwd: GAT GGA GTG TCG GAA CTC ACC (1X)
 
*4EBP1- Rev: GAC CTG GAC AGG ACT CAC CGC (2X)
 
*4EBP1- Rev: GAC CTG GAC AGG ACT CAC CGC (2X)
 
*4EBP1/2- Neo Fwd: GCA TCG AGC GAG CAC GTA CTC (1X)
 
*4EBP1/2- Neo Fwd: GCA TCG AGC GAG CAC GTA CTC (1X)
  
"'4EBP2"'
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WT= 175bp  KO= 550bp
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'''4EBP2'''
 
*4EBP2-Fwd: GGT GGG ACT GTC GGT CTT CTG (1X)
 
*4EBP2-Fwd: GGT GGG ACT GTC GGT CTT CTG (1X)
 
*4EBP2-Rev: CAG CAC CTG GTC ATA GCC GTG (2X)
 
*4EBP2-Rev: CAG CAC CTG GTC ATA GCC GTG (2X)
 
*4EBP1/2- Neo Fwd: GCA TCG AGC GAG CAC GTA CTC (1X)
 
*4EBP1/2- Neo Fwd: GCA TCG AGC GAG CAC GTA CTC (1X)
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 +
WT= 450bp  KO= 750bp
  
 
Make primers at 1uM (Fwd1 + Rev1, Neo Fwd1/2 + Rev1, Fwd2 + Rev2, and Neo Fwd1/2 + Rev2). PCR 4EBP1 and 4EBP2 primers separately.
 
Make primers at 1uM (Fwd1 + Rev1, Neo Fwd1/2 + Rev1, Fwd2 + Rev2, and Neo Fwd1/2 + Rev2). PCR 4EBP1 and 4EBP2 primers separately.

Latest revision as of 14:41, 8 June 2020

PRIMERS

4EBP1

  • 4EBP1- Fwd: GAT GGA GTG TCG GAA CTC ACC (1X)
  • 4EBP1- Rev: GAC CTG GAC AGG ACT CAC CGC (2X)
  • 4EBP1/2- Neo Fwd: GCA TCG AGC GAG CAC GTA CTC (1X)

WT= 175bp KO= 550bp

4EBP2

  • 4EBP2-Fwd: GGT GGG ACT GTC GGT CTT CTG (1X)
  • 4EBP2-Rev: CAG CAC CTG GTC ATA GCC GTG (2X)
  • 4EBP1/2- Neo Fwd: GCA TCG AGC GAG CAC GTA CTC (1X)

WT= 450bp KO= 750bp

Make primers at 1uM (Fwd1 + Rev1, Neo Fwd1/2 + Rev1, Fwd2 + Rev2, and Neo Fwd1/2 + Rev2). PCR 4EBP1 and 4EBP2 primers separately.