IGEM:Groningen/Notebook/iGEM 2011/2011/06/14: Difference between revisions
Joyce Mulder (talk | contribs) (Autocreate 2011/06/14 Entry for IGEM:Groningen/Notebook/iGEM_2011) |
Joyce Mulder (talk | contribs) |
||
Line 6: | Line 6: | ||
| colspan="2"| | | colspan="2"| | ||
<!-- ##### 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. ##### --> | ||
== | ==14-6-11== | ||
<br> | |||
<br> Checking transformants/ testing whether DNA purification from gel went well and is functional in cloning: | |||
<br> PlasB from gel+ vector from gel: no colonies, maybe 1 or 2 | |||
<br> PlasB from gel+ vector no gel: Lots of colonies, might be a lot of self ligation, but the self ligation control is negative... | |||
<br> PBADaraC from gel+ vector from gel: no colonies, just a few but it looks like contamination? | |||
<br> PBADaraC no gel+ vector gel: much colonies | |||
<br> PBADaraC gel + vector no gel: lots of colonies | |||
<br> | |||
<br> Grow colonies , 3 colonies per different construct for plasmid prepping | |||
<br> | |||
<br> Colony PCR | |||
<br> Taq 10× buffer: 40μl | |||
<br> dNTPs 10mM: 8μl | |||
<br> Forward Biobrick primer: 8μl | |||
<br> Reverse Biobrick primer: 8μl | |||
<br> Taq DNA polymerase: 2μl | |||
<br> MQ water: 334μl | |||
<br> | |||
<br> PCR conditions: | |||
<br> Preheated lid: 111°C | |||
<br> Denaturation: 94°C for 10 min. | |||
<br> Cycle 33×: | |||
<br> denaturation: 94°C for 30s. | |||
<br> annealing: 60°C for 30s. | |||
<br> Extension: 72°C for 2,5 min. | |||
<br> Final extension: 72°C for 10 min. | |||
<br> Store infinite at 4°C | |||
<br> | |||
<br> Analyse on a 1% TBE agarose gel. | |||
<br> | |||
<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### --> | <!-- ##### DO NOT edit below this line unless you know what you are doing. ##### --> |
Revision as of 02:38, 21 September 2011
Project name | <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> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> |
14-6-11
|