User:Elaine Marie Robbins/Notebook/CHEM-496/2011/09/21: Difference between revisions

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|style="background-color: #EEE"|[[Image:BDLlogo_notext_lr.png|128px]]<span style="font-size:22px;"> Gel Electrophoresis of GFP</span>
|style="background-color: #EEE"|[[Image:BDLlogo_notext_lr.png|128px]]<span style="font-size:22px;"> Gel Electrophoresis of PCR Product</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"|<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>}}
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==Objective==
==Objective==


To determine whether or not GFP is present in solutions made on [http://openwetware.org/wiki/User:Elaine_Marie_Robbins/Notebook/CHEM-496/2011/09/20| 09/20/11] using agarose gel electrophoresis.
To determine whether or not the PCR performed on [http://openwetware.org/wiki/User:Elaine_Marie_Robbins/Notebook/CHEM-496/2011/09/20 09/20/11] of the DNA coding for a mutant GFP was successful using agarose gel electrophoresis.


==Description==
==Description==
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# Prepare a ladder containing DNA, glycerol, and gel loading dye.
# Prepare a ladder containing DNA, glycerol, and gel loading dye.
# Load DNA solutions and run the gel at 80 V for 40 min.
# Load DNA solutions and run the gel at 80 V for 40 min.
# Remove gel and place in TAE and ethidium bromide (EtBr) solution, mix for 15 min.
# Rinse gel with TAE for 5 min.


==Data==
==Data==


A photo of the electrophoresis gel is displayed below:


<center>[[image:DNA gel 092111 003.jpg]]</center>


The first column is the ladder. The next columns are groups 1, 2, 3, 4, and 5.


==Notes==
==Notes==
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Gel ladder:
Gel ladder:
500 bp, 1000 bp, 2000 bp, 3000 bp, 4000 bp, 5000 bp, 6000 bp, 7000 bp, 8000 bp, 9000 bp, 10000 bp
500 bp, 1000 bp, 2000 bp, 3000 bp, 4000 bp, 5000 bp, 6000 bp, 7000 bp, 8000 bp, 9000 bp, 10000 bp


An optimum gel would have used 1.5% agarose.
An optimum gel would have used 1.5% agarose.

Revision as of 12:07, 4 October 2011

Gel Electrophoresis of PCR Product <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>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>


Objective

To determine whether or not the PCR performed on 09/20/11 of the DNA coding for a mutant GFP was successful using agarose gel electrophoresis.

Description

  1. Make a 1% agarose gel by adding 0.25 g of 0.01 g/mL of agarose to 25 mL of 1x tris base, acetic acid, and ethylenediaminetetraacetic acid (TAE) buffer and heating the mixture in the microwave for 40 s.
  2. Mix 1 μL of 6x gel loading dye with 5 μL of the DNA sample.
  3. Prepare a ladder containing DNA, glycerol, and gel loading dye.
  4. Load DNA solutions and run the gel at 80 V for 40 min.
  5. Remove gel and place in TAE and ethidium bromide (EtBr) solution, mix for 15 min.
  6. Rinse gel with TAE for 5 min.

Data

A photo of the electrophoresis gel is displayed below:

The first column is the ladder. The next columns are groups 1, 2, 3, 4, and 5.

Notes

Gel ladder: 500 bp, 1000 bp, 2000 bp, 3000 bp, 4000 bp, 5000 bp, 6000 bp, 7000 bp, 8000 bp, 9000 bp, 10000 bp

An optimum gel would have used 1.5% agarose.