PCR of Gmab Heavy
Gel shows successful PCR of Gmab Heavy from gBlock mIgM Gmab Heavy.
Forward primer: gBlock F
Reverse primer: gBlock R
Annealing temperature: ?
Run using samples after PCR purification. Extra bands indicated multiple PCR products, used gel extraction to purify out the appropriate band.
There were multiple bands in the gel, indicating multiple PCR products. We used gel extraction to get out the appropriate band.
Restriction Digest Verification of pENTR
7 July 2014:
See gel here
Correct plasmids | Failed plasmids |
|---|---|
Gmab Heavy (1) Gmab Heavy (2) Gmab Heavy (3) Gmab Heavy (4) |
|
Sequencing verification:
| Plasmid | Mutations | Where? |
|---|---|---|
| Gmab Heavy (1) | G45 -> V | two variable, |
| Gmab Heavy (2) | two nonsense | no |
| Gmab Heavy (3) | G207 -> W | both constant region |
| Gmab Heavy (4) | K84 -> M | variable region |
| Gmab Heavy (11) | Y82 -> H | variable region |
| Gmab Heavy (12) | no alignment | |
| Gmab Heavy (13) | P290 -> T | constant region |
| Gmab Heavy (14) | none | worked |
| Gmab Heavy (15) | no alignment | |
| Gmab Heavy (16) | one nonsense | no |
| Gmab Heavy (17) | none | worked |
| Gmab Heavy (18) | none | worked |
| Gmab Heavy (19) | one nonsense, two frameshift | no |
| Gmab Heavy (20) | none | worked |
| Gmab Heavy (21) | no alignment | |
| Gmab Heavy (22) | no alignment |