IGEM:Slovenia HS 2015/2009/Notebook/Test/2015/07/21: Difference between revisions
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== | ==CtfA, CtfB, BdhB restriction== | ||
* | Mixture: | ||
* 20 μL DNA | |||
* 2,5 μL buffer Tango | |||
* 1 μL Xba1 | |||
* 2 μL Psrt1 | |||
Restriction mixture was incubated at 37°C for 3h. | |||
==PCR== | |||
* PCR reaction was conducted accordingly to [http://openwetware.org/wiki/FCCT_Biochemistry_Lab:PCR_protocol_iGEM_HS PCR]<br> | |||
* [http://openwetware.org/wiki/FCCT_Biochemistry_Lab: PCR Master Mix]<br> | |||
Markings: | |||
1-10 CtfA, | |||
11-20 CtfB, | |||
21-30 BdhB | |||
==Agarose gel electrophoresis== | |||
*[http://openwetware.org/wiki/FCCT_Biochemistry_Lab:Agarose_gel_electrophoresis Agarose gel electrophoresis]<br> | |||
* We have performed two electrophoresis ( for PCR products, restriction products) | |||
Expected lenghts of genes: | |||
* CtfA: 735 bp | |||
* CtfB: 744 bp | |||
* BdhA: 1250 bp | |||
* AdhE1: 1666 bp | |||
* AdhE2: 1020 BP | |||
Electrophoresis was successful for the restriction products. Fragments with appropriate lenghts were cut from the gel. | |||
==DNA isolation from agarose gel== | |||
*protocol- manjka še! | |||
==Ligation== | |||
po protokolu- manjka še! | |||
* overnight ligation | |||
These fragments were then inserted into the expression vector pSB1C3. | |||
Revision as of 03:55, 26 July 2015
 iGEM Project name 1
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CtfA, CtfB, BdhB restrictionMixture:
Restriction mixture was incubated at 37°C for 3h. PCR
Markings: 1-10 CtfA, 11-20 CtfB, 21-30 BdhB Agarose gel electrophoresis
Expected lenghts of genes:
Electrophoresis was successful for the restriction products. Fragments with appropriate lenghts were cut from the gel. DNA isolation from agarose gel
Ligationpo protokolu- manjka še!
These fragments were then inserted into the expression vector pSB1C3.
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