User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/11/06: Difference between revisions
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* Cell culture was taken out of shaker, and plated on LB agar plates made with LB, agar, and ampicillin. The LB plates were made in the following way: | * Cell culture was taken out of shaker, and plated on LB agar plates made with LB, agar, and ampicillin. The LB plates were made in the following way: | ||
Following reagents were weighted out: | |||
20g of LB Agar powder from Fisher Scientific® | |||
5g of tryptone from Fisher Scientific® | |||
2.5g of yeast extract from Fisher Scientific® | |||
5g of NaCl from Fisher Scientific® | |||
7.5g agar from Fisher Scientific® | |||
Above were placed into a 800mL Erlenmeyer flask. | |||
500mL distilled water was added to reagents. | |||
The solution was autoclaved under liquid cycle (see [[User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/11/03|2012/11/03]] for protocol for liquid cycle) | |||
After cooling to ~50°C, 50mg ampicillin was poured into autoclaved solution, swirled to mix | |||
Under sterile condition, the solution was poured into separate petri dish when temperature reaches ~40°C | |||
Plates were left under sterile condition to solidify | |||
* 2 plates were yielded: one with 50μL of transformed cell culture, the other with 200μL of transformed cell culture. | |||
* Plates were placed in incubator for 15 hours upside down for cellular growth. | |||
==Results for E.coli cell Transformation== | ==Results for E.coli cell Transformation== |
Revision as of 21:08, 24 November 2012
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Purpose
Procedure for Protein Extraction and Purification from BL21(DE3) Cells
Speed: 18,000rpm Temperature: 4°C Time elapsed: 2 hours Rotor used:_____
Result for Protein Extraction and Purification from BL21(DE3) Cells
Procedure for E.coli cell Transformation
225rpm 37°C 1 hour
Following reagents were weighted out: 20g of LB Agar powder from Fisher Scientific® 5g of tryptone from Fisher Scientific® 2.5g of yeast extract from Fisher Scientific® 5g of NaCl from Fisher Scientific® 7.5g agar from Fisher Scientific® Above were placed into a 800mL Erlenmeyer flask. 500mL distilled water was added to reagents. The solution was autoclaved under liquid cycle (see 2012/11/03 for protocol for liquid cycle) After cooling to ~50°C, 50mg ampicillin was poured into autoclaved solution, swirled to mix Under sterile condition, the solution was poured into separate petri dish when temperature reaches ~40°C Plates were left under sterile condition to solidify
Results for E.coli cell Transformation
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