User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2013/02/05

Current revision

Experimental Biological Chemistry II

Procedure for grinding of Ag+ Clay into powder form is recorded in Melissa's Notebook.
Growing of DH5α-T1 cells
- One shot of DH5α-T1 cells was taken out from -87°C fridge.
- Cells were thawed in room temperature
- 3mL of autoclaved LB media was pipetted into a test tube
- Inoculation loop was used to obtain a small portion of cells. The cells were then dipped into the 3mL LB media.
- The cell culture was then placed in orbital shaker for 2 hours, at 37°C, with shaking at 230rpm.
- After two hours, the culture appeared clear, indicating no cell growth, an alternative approach were taken.
A different cell line was used for cell amplification, and the procedure for DH5α-T1 cell amplification was altered then repeated.
- DH10B cells were taken from -87°C fridge.
- The cell were let thaw on ice.
- In two clean 15mL falcon tubes, 3mL autoclaved LB media were added.
- In one falcon tube, 10uL DH10B cells were added.
- In another falcon tube, 10uL DH5α-T1 cells were added.
- The two falcon tubes were placed in orbital shaker for 2 hours, at 37°C, with shaking at 230rpm.
- After 2 hours of incubation, the both cultures appeared clear. As a result, both cultures were placed in incubator horizontally to maximize shaking and left in shaker under same conditions for over night.

@@ Line 18: / Line 18: @@
 ** Inoculation loop was used to obtain a small portion of cells. The cells were then dipped into the 3mL LB media.
 ** The cell culture was then placed in orbital shaker for 2 hours, at 37°C, with shaking at 230rpm.
+** After two hours, the culture appeared clear, indicating no cell growth, an alternative approach were taken.
+* A different cell line was used for cell amplification, and the procedure for DH5α-T1 cell amplification was altered then repeated.
+** DH10B cells were taken from -87°C fridge.
+** The cell were let thaw on ice.
+** In two clean 15mL falcon tubes, 3mL autoclaved LB media were added.
+** In one falcon tube, 10uL DH10B cells were added.
+** In another falcon tube, 10uL DH5α-T1 cells were added.
+** The two falcon tubes were placed in orbital shaker for 2 hours, at 37°C, with shaking at 230rpm.
+** After 2 hours of incubation, the both cultures appeared clear. As a result, both cultures were placed in incubator horizontally to maximize shaking and left in shaker under same conditions for over night.