User:Puja Mody/Notebook/Chem 571: Gold Nanoparticles/2012/10/24: Difference between revisions
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|style="background-color: #F2F2F2" align="center"| | |style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} | ||
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==Procedure== | ==Procedure== | ||
* The PCR product was transformed by following the [[AU Biomaterials Design Lab:Protocols/Transformation Protocol| Transformation Protocol]] | * The PCR product was transformed by following the [[AU Biomaterials Design Lab:Protocols/Transformation Protocol| Transformation Protocol]] | ||
*The two solutions of 1μL of DPN1 was added to 45μL of K110A (f) and K110A (r) plasmid that had been stored in the freezer were used for cell transformation. | |||
* 1 vial of the cells was thawed on ice. 10ng of DNA in 5μL was added to 40 microliters of cells which were then heat shocked by incubating in a 42°C water bath for 30 seconds. | |||
The | *Upon removal, the vial was placed immediately on ice. | ||
*200μL of SOC media was added to one solution of cells while 200 μL of LB was added to the other solution of cells. Both vials were shaken at 275 rpm for 1 hour. | |||
*Next, 25 μL of transformed cells were plated on one LB plate petridish (which was prewarmed and contained LB and kanamycin) while 200 μL of transformed cells was placed on the other. *The plates were incubated at 37°C overnight. | |||
* ADA protein was expressed in the same manner as on [[User:Puja Mody/Notebook/Chem 571: Gold Nanoparticles/2012/09/19| 2012/09/19]] | * ADA protein was expressed in the same manner as on [[User:Puja Mody/Notebook/Chem 571: Gold Nanoparticles/2012/09/19| 2012/09/19]] |
Latest revision as of 22:08, 26 September 2017
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Transformation, protein expression, lysozyme Uv-VisGoals
Procedure
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