Difference between revisions of "4EBP1/2 Genotyping"
From Bridges Lab Protocols
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==PRIMERS== | ==PRIMERS== | ||
| − | + | '''4EBP1''' | |
| − | + | ||
*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) | ||
| − | + | WT= 175bp KO= 550bp | |
| + | |||
| + | '''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) | ||
| + | |||
| + | 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.
