Team:SDU-Denmark/labwork/Notebook/week4

From 2012.igem.org

(Difference between revisions)
 
(One intermediate revision not shown)
Line 298: Line 298:
<p> <b>23-07-2012 to 29-07-2012</b> </p>
<p> <b>23-07-2012 to 29-07-2012</b> </p>
-
<h2>SST send for sequencing </h2>
 
<p>
<p>
 +
Another checkdigest was made on SST colonies, we had our doubts about the entegrity of the formerly used restriction enzyme, so a newer EcoRI was used, a get was made and this time we got results. After a checkdigest that showed bands at expected lengths and nanodrop, two samples of 1-SST was sent for sequencing.<br/><br/>
-
SST from the gel extraction the 21-07-2012 was diluted to a concentration of 100ng/µL and separated to 4 samples of 15µL each and send of for sequencing. <br/>
+
4 days later we recieved results from sequencing. As we expected from the original ncbi sequences, the genes contained illegal restriction sites.<br/>
-
We made liquid cultures from the SST plates from 19-07-2012 and incubated them O.N.
+
1-SST contained 1 SpeI restriction site and 1-FFT had 1 EcoRI and 1 XbaI. <br/><br/>
-
 
+
-
<br/> 
+
-
<br/> 
+
-
 
+
-
<h2>Cryo backup, miniprep and preparation to sequencing </h2>
+
-
<p>
+
-
 
+
-
We made some Cryo backup’s from the 18 liquid cultures, that was incubated O.N. and took some sample out from each to do a miniprep on. <br/>  
+
-
We ran a gel on the samples and found that two samples had bands corresponding to our cut gene and vector.  
+
-
These will be shipped of to be sequneced.
+
-
+
-
<br/> <br/>
+
-
 
+
-
<h2> SST send for sequencing </h2>
+
-
<p>
+
-
 
+
-
The two sample from earlier were prepared for sequencing by diluting them to 70-78 ng/μl and then send. Afterwards we looked into the expected outcomes of sequencing and had a look at the following mutagenesis post-sequencing.
+
-
 
+
 +
Primers for mutagenesis were constructed and ordered with the help of our supervisor Steffen Schmidt
 +
</p>

Latest revision as of 02:07, 27 September 2012

iGEM TEAM ::: SDU-DENMARK

Laboratory Notebook

1st week 2nd week 3th week 4th week
5th week 6th week 7th week 8th week
9th week 10th week 11th week 12th week

23-07-2012 to 29-07-2012

Another checkdigest was made on SST colonies, we had our doubts about the entegrity of the formerly used restriction enzyme, so a newer EcoRI was used, a get was made and this time we got results. After a checkdigest that showed bands at expected lengths and nanodrop, two samples of 1-SST was sent for sequencing.

4 days later we recieved results from sequencing. As we expected from the original ncbi sequences, the genes contained illegal restriction sites.
1-SST contained 1 SpeI restriction site and 1-FFT had 1 EcoRI and 1 XbaI.

Primers for mutagenesis were constructed and ordered with the help of our supervisor Steffen Schmidt

Retrieved from "http://2012.igem.org/Team:SDU-Denmark/labwork/Notebook/week4"