840:153g:Projects/project22/2012/10/18

From OpenWetWare

< 840:153g:Projects | project22 | 2012 | 10(Difference between revisions)
Jump to: navigation, search
(PCR and Gel Electrophoresis Check)
Current revision (16:40, 25 October 2012) (view source)
(PCR and Gel Electrophoresis Check)
 
(2 intermediate revisions not shown.)
Line 7: Line 7:
<!-- ##### 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. ##### -->
==PCR and Gel Electrophoresis Check==
==PCR and Gel Electrophoresis Check==
-
On Tuesday we decided that even though the gel electrophoresis was flawed...
+
On Tuesday we decided that even though the gel electrophoresis was flawed due to anomalous specks on the gel...
[[Image:Small_gel_10.11.12.png‎]] <-- notice anomalous specks on gel
[[Image:Small_gel_10.11.12.png‎]] <-- notice anomalous specks on gel
Line 17: Line 17:
[[Image:Small_gel_10.18.12.png]]
[[Image:Small_gel_10.18.12.png]]
-
As you can see, our positive control worked (4th lane from the left), but our samples did not display the expected brightness. It is important to note that our annealing time was only 1 minute 30 seconds and our target gene is 1470 b.p., so that may have an effect on our outcome. It is also possible our DNA is compromised and we will need to perform another DNA extraction.
+
As you can see, our positive control worked (4th lane from the left), but our samples (lanes 5 and 6 from the left) did not display the expected brightness. It is important to note that our annealing time was only 1 minute 30 seconds and our target gene is 1470 b.p., so that may have an effect on our outcome. It is also possible our DNA is compromised (and we will need to perform another DNA extraction) or our primers are flawed (aren't working properly).
For next Tuesday, we will increase the annealing time for PCR and see if we get better results. We will also grow up more cultures of ''S. marcescens'' and prepare for DNA extraction with alternate protocols obtained from Open WetWare (the last ones we used were obtained from the USDA).
For next Tuesday, we will increase the annealing time for PCR and see if we get better results. We will also grow up more cultures of ''S. marcescens'' and prepare for DNA extraction with alternate protocols obtained from Open WetWare (the last ones we used were obtained from the USDA).

Current revision

Prodigiosin Production in E.Coli Main project page
Previous entry      Next entry

PCR and Gel Electrophoresis Check

On Tuesday we decided that even though the gel electrophoresis was flawed due to anomalous specks on the gel...

Image:Small_gel_10.11.12.png‎ <-- notice anomalous specks on gel

...we felt we had sufficient DNA to proceed to PCR. Our purpose was two-fold - to make sure the positive control worked (last time we ran PCR it did not) and to see if our DNA was truly present in the sample (PCR would not amplify the gene if DNA wasn't there).

On Thursday, we ran gel electrophoresis to check our PCR.

Image:Small_gel_10.18.12.png

As you can see, our positive control worked (4th lane from the left), but our samples (lanes 5 and 6 from the left) did not display the expected brightness. It is important to note that our annealing time was only 1 minute 30 seconds and our target gene is 1470 b.p., so that may have an effect on our outcome. It is also possible our DNA is compromised (and we will need to perform another DNA extraction) or our primers are flawed (aren't working properly).

For next Tuesday, we will increase the annealing time for PCR and see if we get better results. We will also grow up more cultures of S. marcescens and prepare for DNA extraction with alternate protocols obtained from Open WetWare (the last ones we used were obtained from the USDA).


Personal tools