User:Laura Flynn/Notebook/Experimental Biological Chemistry/2011/11/02: Difference between revisions
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==Objective== | ==Objective== | ||
To create AUNP fibers using BSA and HAUCl<sub>4</sub>. | To load and run mutated DNA on gel electrophoresis | ||
To create and autoclave an LB Agar solution | |||
To create AUNP fibers using BSA and HAUCl<sub>4</sub>. | |||
==Procedure== | ==Procedure== | ||
AuNP Sythesis: | |||
New HAuCl<sub>4</sub> stock was made. Protocol was followed from the previous week. Three test tubes were used: | New HAuCl<sub>4</sub> stock was made. Protocol was followed from the previous week. Three test tubes were used: | ||
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The oven was kept at approximately 70 degrees. | The oven was kept at approximately 70 degrees. | ||
DNA Transformation: | |||
*Non-methylated DNA was digested with 1 μL Dpnl. | |||
*LB/agar plates were made using: | |||
**0.875 g LB, 0.7 g agar, and 35 mL of water. | |||
*The mixture was then autoclaved. | |||
* 35 μL ampicillin was added before the agar completely cooled. | |||
*The agar was then poured onto the plate and left to solidify. | |||
* Both a sterile tube and the desired DNA were placed in an ice bucket for 15 minutes. | |||
* 5 μL of DNA and 30 μL of cells were added to the bottom of the sterile tube, then incubated on ice for 30 minutes. | |||
*The DNA/cells were then heat shocked at 42°C for 30 s, and incubated on ice for 5 minutes. | |||
* 250 μL of SOC media was added to the tube, which was then incubated in the shaker at 37°C for 1 hr. | |||
*100 μL of the cells were spread on the LB/agar plate using sterile technique. | |||
* The plate was then stored inverted overnight in an oven at 37°C oven. | |||
Electrophoresis: | |||
* a 1% agarose gel was made by adding 0.25 g of a 0.01 g/mL agarose solution to 25 mL of a 1x tris base, acetic acid, and TAE buffer. This mixture was then microwaved for 40 seconds. | |||
* 2 μL of 6x gel loading dye was mixed with 10 μL of the DNA sample. | |||
* A ladder was prepared containing DNA, glycerol, and gel loading dye. | |||
* Each DNA solution was loaded and the gel was run at 80 V for 40 minutes. | |||
* The gel was removed and placed in TAE and EtBr solution, then mixed for 15 minutes. | |||
* The gel was rinsed for 5 minutes with TAE. | |||
==Results== | ==Results== | ||
Each test tube formed purple fibers. Test tube one's fibers were long, while the other two test tubes had shorts more aggregated fiber clumps. | |||
==Notes== | ==Notes== |
Revision as of 13:21, 30 November 2011
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ObjectiveTo load and run mutated DNA on gel electrophoresis To create and autoclave an LB Agar solution To create AUNP fibers using BSA and HAUCl4. ProcedureAuNP Sythesis: New HAuCl4 stock was made. Protocol was followed from the previous week. Three test tubes were used: Each test tube contained 1 mL of BSA, 1 mL of HAuCl4, and 8 mL of water.
The oven was kept at approximately 70 degrees. DNA Transformation:
Electrophoresis:
ResultsEach test tube formed purple fibers. Test tube one's fibers were long, while the other two test tubes had shorts more aggregated fiber clumps. NotesAuNP:
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