User:Trisha I. Ibeh/Notebook/Trisha Notebook/2013/09/04: Difference between revisions

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|style="background-color: #EEE"|[[Image:BDLlogo_notext_lr.png|128px]]<span style="font-size:22px;"> Biomaterials Design Lab</span>
|style="background-color: #EEE"|[[Image:BDLlogo_notext_lr.png|128px]]<span style="font-size:22px;"> Biomaterials Design Lab</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>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</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"|[[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]]&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}}
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The same procedure that was performed with adenosine will be performed with inosine. The procedure can be found [[User:Matt_Hartings/Notebook/AU_Biomaterials_Design_Lab/2013/09/03|here.]]
The same procedure that was performed with adenosine will be performed with inosine. The procedure can be found [[User:Matt_Hartings/Notebook/AU_Biomaterials_Design_Lab/2013/09/03|here.]]


==Instructions==
==Data==
Each group will pick a different protein (bovine serum albumin, horseradish peroxidase, pepsin, adenosine deaminase, hemoglobin). You'll need to make 4 samples ranging from 10uM to 0.5uM. Remember, we don't have a lot of most of these proteins, so figure out how to make a stock solution using as little solid protein as possible. The fluorescence cuvette needs only 200uL, so you don't need to make more than that for each concentration that you're measuring.
'''Inosine Abs Spectra'''


# Make your solutions
[[Image:Screen_Shot_2013-10-05_at_6.27.27_AM.png]]<br>
# Take an absorbance measurement of your samples and a blank
# Take a fluorescence measurement of your samples and a blank (excitation: 290nm, emission: 310nm-500nm)
# Analyze your data. (Integrate your corrected spectrum. Integration is MUCH more important in fluorescence spectra than peak height.) [http://people.oregonstate.edu/~haggertr/487/integrate.htm Here's] a site from the web that talks about how to integrate using formulas in excel.
# Normalize your corrected spectra to the absorption intensity at 280nm for the corresponding absorbance spectrum. (That is, divide the fluorescence intensity by the A(280) and plot all of the fluorescence spectra on the same chart. Do they match up? Are they different? What could this mean?)


**Note --- We are only going to have 1 cuvette to use. I expect you to work quickly and spend time analyzing data.
'''Inosine Calibration Curve'''
 
[[Image: Screen_Shot_2013-10-05_at_6.13.30_AM.png]]<br>
 
'''Class Data (pooled)'''
 
[[Image: Screen_Shot_2013-10-05_at_6.57.56_AM.png]] <br>
 
[[Image: Screen_Shot_2013-10-05_at_6.31.40_AM.png]]<br>
 
'''Abs mean and std dev (adenosine and inosine)'''
 
'''Absorbance Spectra of Unknown'''
 
[[Image: Screen_Shot_2013-10-05_at_7.07.13_AM.png]]<br>


==Notes==
==Notes==

Latest revision as of 23:16, 26 September 2017

Biomaterials Design Lab Main project page
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Objective

Finish up the lab from the previous day (9/3/13) by taking the UV- Vis spectrum of inosine and doing data analysis. From the data, a Grubb's test will be performed to determine the outliers. The concentration of the unknown sample will be determined ad the molar absorbtivity will also be determined from the class data.

Procedure

The same procedure that was performed with adenosine will be performed with inosine. The procedure can be found here.

Data

Inosine Abs Spectra


Inosine Calibration Curve


Class Data (pooled)



Abs mean and std dev (adenosine and inosine)

Absorbance Spectra of Unknown


Notes

I made a stock solution of BSA today. Here is the info:

BSA solution 0.0104g BSA (MW = 66776g/mol) in 0.0100mL water → 15.6μM




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