User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2013/02/27: Difference between revisions
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Experimental Biological Chemistry II</span> | ||
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html> </html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}} | |style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html> </html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}} | ||
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<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### --> | <!-- ##### DO NOT edit above this line unless you know what you are doing. ##### --> | ||
== | ==Purpose== | ||
* | * Run pilot study on cell growth with concentrations of AgNO<sub>3</sub> in solution varying from 1uM to 10mM | ||
==Procedure== | |||
* DH5α-T1 cell pellets were taken out from -80°C freezer and left in room temperature to defrost. | |||
* Under a sterile environment, 3mL of autoclaved LB medium were added onto pellet. The solution was pipetted up and down gently to mix. | |||
* In five bottles of 125mL Erlenmeyer flasks containing 50mL LB medium, the following amount of LB were taken out of the medium from each flask: | |||
4.76uL, 5.00uL, 50.0uL, 500uL, 5000uL | |||
* Respectively, the following amount and concentrations of AgNO<sub>3</sub> were placed in: | |||
{| {{table}} | |||
| align="center" style="background:#f0f0f0;"|'''Volume of LB taken out(uL)''' | |||
| align="center" style="background:#f0f0f0;"|'''Volume of AgNO3 added(uL)''' | |||
| align="center" style="background:#f0f0f0;"|'''Concentration of AgNO3 added(M)''' | |||
| align="center" style="background:#f0f0f0;"|'''Final Concentration of AgNO3 in flask(uM)''' | |||
|- | |||
| 4.76||4.76||0.0105||1 | |||
|- | |||
| 5||5||0.1||10 | |||
|- | |||
| 50||50||0.1||100 | |||
|- | |||
| 500||500||0.1||1,000 | |||
|- | |||
| 5000||5000||0.1||10,000 | |||
|} | |||
* Then, 1mL of resuspended cells were placed into each of the five flasks. | |||
* The positive control contains 50mL of LB media and 1mL of cells | |||
* One more control was set up containing 45mL of LB media and 5mL of 0.1M AgNO<sub>3</sub>. | |||
* All seven flasks were placed into orbital shaker at 230rpm, at 37°C, for 15 hour time period. | |||
* Absorbance on a dual-beam spectrometer was set up under photometric method, wavelength at 600nm, to measure the absorbance of each cell solution at each hour. | |||
* 3mL of LB solution was used as blank for all measurements, and 3mL of sample was taken each time for absorbance readings. | |||
==Results== | |||
* Seven samples were run for a period of 15 hours, and absorbance at 600nm was taken at every hour. | |||
* The absorbance for each sample at each hours were shown in the following table: | |||
{| {{table}} | |||
| align="center" style="background:#f0f0f0;"|'''Hour''' | |||
| align="center" style="background:#f0f0f0;"|'''Control#1 (LB+Cell)''' | |||
| align="center" style="background:#f0f0f0;"|'''Control#2 (LB+5mL AgNO3)''' | |||
| align="center" style="background:#f0f0f0;"|'''1uM AgNO3 + Cell''' | |||
| align="center" style="background:#f0f0f0;"|'''10uM AGNO3 + cell''' | |||
| align="center" style="background:#f0f0f0;"|'''100uM AGNO3 + cell''' | |||
| align="center" style="background:#f0f0f0;"|'''1mM AGNO3 + cell''' | |||
| align="center" style="background:#f0f0f0;"|'''10mM AGNO3 + cell''' | |||
|- | |||
| 0||0.113||0.366||0.116||0.122||0.148||0.178||0.387 | |||
|- | |||
| 1||0.179||0.491||0.181||0.148||0.143||0.185||0.516 | |||
|- | |||
| 2||0.417||0.583||0.438||0.247||0.138||0.189||0.595 | |||
|- | |||
| 3||0.77||0.682||0.784||0.515||0.136||0.2||0.674 | |||
|- | |||
| 4||1.296||0.783||1.258||0.925||0.134||0.201||0.754 | |||
|- | |||
| 5||1.638||0.859||1.383||1.628||0.134||0.202||0.815 | |||
|- | |||
| 6||1.779||0.943||1.771||1.647||0.134||0.208||0.877 | |||
|- | |||
| 7||1.873||1.031||1.865||1.793||0.13||0.209||0.942 | |||
|- | |||
| 8||1.929||1.098||1.933||1.872||0.132||0.213||0.987 | |||
|- | |||
| 9||1.975||1.174||1.971||1.92||0.131||0.214||1.044 | |||
|- | |||
| 10||2.016||1.247||2.005||1.953||0.131||0.217||1.096 | |||
|- | |||
| 11||2.055||1.303||2.033||1.992||0.132||0.222||1.152 | |||
|- | |||
| 12||2.06||1.37||2.06||2.008||0.132||0.225||1.201 | |||
|- | |||
| 13||2.062||1.429||2.055||2.008||0.131||0.228||1.248 | |||
|- | |||
| 14||2.068||1.486||2.052||2.007||0.128||0.23||1.3 | |||
|- | |||
| 15||2.068||1.544||2.053||2.007||0.13||0.233||1.34 | |||
|} | |||
* The graph Absorbance versus Cell growth varying concetration of AgNO<sub>3</sub> is plotted in graph below: | |||
[[Image:Absorbance versus Cell growth in different concentrations of AgNO3.jpg|600px]] | |||
==Observation== | |||
* After the addition of 0.1M AgNO<sub>3</sub> into 50mL LB medium, the solution immediately turned opaque. Due to the small amount of AgNO<sub>3</sub> added into solutions containing AgNO<sub>3</sub> with the following concentration: 1uM, 10uM, 100uM, 1mM, and due to the limit of LB medium present, no controls were set for these samples. | |||
* The solution containing cells and 10mM AgNO<sub>3</sub> appeared to be the most opaque, a control that contained 45mL LB and 5mL 0.1M AgNO<sub>3</sub> were also run at the same time. | |||
* The opaque color of solution containing cells 10mM AgNO<sub>3</sub>, as well as the control containing 45mL LB and 5mL of 0.1M AgNO<sub>3</sub>, turned darker over time, eventually, at the point of 15 hours, the color of solution turned darkish gray. | |||
* The opaque color might be due to silver ions in solution being oxidized by the chloride present in LB media, turning a darker color. | |||
Revision as of 21:28, 27 February 2013
Experimental Biological Chemistry II | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Purpose
Procedure
4.76uL, 5.00uL, 50.0uL, 500uL, 5000uL
Results
Observation
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