User:Puja Mody/Notebook/Chem 571: Gold Nanoparticles/2012/09/18: Difference between revisions
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span> | ||
|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== | ||
* [[ | *Following the Starter Culture media protocol and the expression culture media protocol, we created LB broth for growing cells. The Starter culture media was used mainly today and was eventually combined with the expression culture media. | ||
* [[AU_Biomaterials_Design_Lab:Protocols/Starter_Culture_Media| Started Culture Protocol]] | |||
** add 35 mL H<sub>2</sub>Oto each to 4 250mL erlenmeyer flask | ** add 35 mL H<sub>2</sub>Oto each to 4 250mL erlenmeyer flask | ||
** [[AU_Biomaterials_Design_Lab:Materials/LB|LB]] was added to each flask. | ** [[AU_Biomaterials_Design_Lab:Materials/LB|LB]] was added to each flask. | ||
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(note that while there were the amounts measured out- the amount of LB added to the flasks was actually less due to loss of sample in transferring it from the weigh dish to the flask) | (note that while there were the amounts measured out- the amount of LB added to the flasks was actually less due to loss of sample in transferring it from the weigh dish to the flask) | ||
* [[ | * [[AU_Biomaterials_Design_Lab:Protocols/Expression_Culture_Media| Expression Culture Media]] | ||
** 1L of distilled water were added to 4.8L Fernbach flasks | ** 1L of distilled water were added to 4.8L Fernbach flasks | ||
**LB was added to each flask | **LB was added to each flask | ||
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* Aseptic techniques were applied by executing the inoculation process near the proximity of a Bunsen burner. The aluminum foil tops of the Erlenmeyer containing the broths were flamed in between uncapping and capping. The metal tweezers were flamed before and after use. Sterile wood sticks were used to capture a colony of the ''E.Coli'' | * Aseptic techniques were applied by executing the inoculation process near the proximity of a Bunsen burner. The aluminum foil tops of the Erlenmeyer containing the broths were flamed in between uncapping and capping. The metal tweezers were flamed before and after use. Sterile wood sticks were used to capture a colony of the ''E.Coli'' | ||
*The starter culture media flasks were then placed in a shaker incubator at 37°C at 237rpm overnight. | |||
Preparation of Tris Buffer | |||
*pH 10- this buffer would be used with the AuNP/BSA made at a concentration of 70 [[User:Puja Mody/Notebook/Chem 571: Gold Nanoparticles/2012/09/12| last week]] | |||
the desired concentration of the stock was .1 M tris and a total of 25mL of stock was made. | |||
1 mol/ 1L x .025 L= .025 mol | |||
.025 mol x 121.14 grams/1 mol = 3.0285 grams of tris in 25mL of water | |||
* in actuality, 3.080 grams of tris was added. | |||
Calculations for amount of water and tris needed to achieve desired concentrations: | |||
{| {{table}} | |||
| align="center" style="background:#f0f0f0;"|'''Concentration of Tris (mM)''' | |||
| align="center" style="background:#f0f0f0;"|'''mL of water''' | |||
| align="center" style="background:#f0f0f0;"|'''Amount of tris to be added (mL)''' | |||
|- | |||
| 0.05||0.994||0.00625 | |||
|- | |||
| 5||0.998||0.0125 | |||
|- | |||
| 5||0.975||0.025 | |||
|- | |||
| 50||0.95||0.05 | |||
|- | |||
| 100||0.9||0.1 | |||
|- | |||
| 200||0.8||0.2 | |||
|- | |||
| 500||0.5||0.5 | |||
|- | |||
| 0.1||0||1 | |||
|} | |||
(The tris and water were not combined with the AuNP/BSa solutions today) | |||
Latest revision as of 22:00, 26 September 2017
Project name | Main project page Previous entry Next entry | |||||||||||||||||||||||||||
Day 5
Goals
Procedure
(note that while there were the amounts measured out- the amount of LB added to the flasks was actually less due to loss of sample in transferring it from the weigh dish to the flask)
Using the formula M1V1=M2V2 The volume or amount of kanamycin that would need to go into each of the flasks was 50 μg/mL x 35mL ÷50,000 μg = .035mL Kanamycin.
Preparation of Tris Buffer
the desired concentration of the stock was .1 M tris and a total of 25mL of stock was made. 1 mol/ 1L x .025 L= .025 mol .025 mol x 121.14 grams/1 mol = 3.0285 grams of tris in 25mL of water * in actuality, 3.080 grams of tris was added.
(The tris and water were not combined with the AuNP/BSa solutions today)
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