840:153g:Projects/project26/2012/11/08: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
No edit summary
Line 1: Line 1:
{|{{table}} width="800"
{|{{table}} width="800"
|-
|-
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Cloning of Atrolysin A</span>
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Cloning of Atrolysin A from ''Crotalus atrox''</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>}}
|-
|-
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 ==
==PCR ==
This week We used our working primers to try and make a PCR product. Our first trial was a fail so we Then added a higher concentration of DNA and lowered our annealing temps these we are hoping allow for a positive DNA results. We also this week isolated our plasmid BBa_I0500 which is 1210 Bp long, from our vector. We ran the DNA on a gel and got to see if the DNA was present or not. And there was faint banding around 1200 bp which is a good sign. If present we will then be able to add our bio brick parts and incorporate our DNA in the plasmid.
This week we used our working primers to try and make a PCR product. Our first trial failed so we then added a higher concentration of DNA and lowered our annealing temps. We are hoping allow for a positive DNA result. We also isolated our plasmid BBa_I0500 which is 1210 Bp long. We ran the sample on a gel and got to see if the DNA was present or not. There was faint banding around 1200 bp which is the size of the insert and banding at 4425 bp which is the size of the backbone. If present we will then be able to add our bio brick parts and incorporate our DNA in the plasmid.


[[Image:11-6-12A.tif]]
[[Image:11-6-12A.tif]]

Revision as of 11:12, 4 December 2012

Cloning of Atrolysin A from Crotalus atrox <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>

PCR

This week we used our working primers to try and make a PCR product. Our first trial failed so we then added a higher concentration of DNA and lowered our annealing temps. We are hoping allow for a positive DNA result. We also isolated our plasmid BBa_I0500 which is 1210 Bp long. We ran the sample on a gel and got to see if the DNA was present or not. There was faint banding around 1200 bp which is the size of the insert and banding at 4425 bp which is the size of the backbone. If present we will then be able to add our bio brick parts and incorporate our DNA in the plasmid.

Starting from the left: there is the positive control, negative control, E3, A3, Ladder, E2, A2, E1, A1. No bands showed up on the gel meaning that there was no amplification.

This gel is of the plasmid DNA. Starting from the left is: K4, K3, Ladder, K2, K1. The bands that showed up were light, but present. They showed up around the 1200 bp mark.