*We have ordered specific oligonucleotides (BdhA, CtfAB).Our goal was the isolation of genes reffering to ''Clostridium acetobutylicum''. There is a table beneath. As oligonucleotides we have used BdhA (R), Bdha (F), ctfAB (R), ctfAB (F), 16S (F), 16S (R). Basically we were changing only the source of isolation.
*We have ordered new specific oligonucleotides (BdhA, CtfAB).Our goal was the isolation of genes reffering to ''Clostridium acetobutylicum''. There is a table beneath. As oligonucleotides we have used BdhA (R), Bdha (F), ctfAB (R), ctfAB (F), 16S (F), 16S (R). Basically we were changing only the source of isolation.
Isolation sources:
Isolation sources:
*broth
*broth
Line 13:
Line 13:
* 100x diluted DNA
* 100x diluted DNA
== Mixture for PCR reaction (master mix) ==
::{| border="1" cellpadding="2" cellspacing="0"
|-
!component||||quantity (µL)||
|-
|oligo 1 (10µM)||1||10||
|-
|oligo 2 (10µM)||1||10||
|-
|dNTP (2mM)||1 ||10||
|-
|MgCl2||2||20||
|-
|buffer||2||20||
|-
|dH2O||11||110||
|-
|Vtot.||/||190||
|-
=Mixture for PCR reaction (master mix)=
Mixture for PCR reaction (Master Mix) was prepared accordingly to [http://openwetware.org/wiki/FCCT_Biochemistry_Lab: PCR Master Mix]
|}
==Electrophoresis==
=Electrophoresis=
*Agarose electrophoresis was the next step.
The next step was Agarose gel electrophoresis. We can see that there are some of the products, but they do not correspond of all the masses of the products. CfAB and BdhA are both 1000 bp long. 16S is 600 bp long. The reaction have to be repeated.
We have ordered new specific oligonucleotides (BdhA, CtfAB).Our goal was the isolation of genes reffering to Clostridium acetobutylicum. There is a table beneath. As oligonucleotides we have used BdhA (R), Bdha (F), ctfAB (R), ctfAB (F), 16S (F), 16S (R). Basically we were changing only the source of isolation.
Isolation sources:
broth
10x diluted broth
DNA
100x diluted DNA
Mixture for PCR reaction (master mix)
Mixture for PCR reaction (Master Mix) was prepared accordingly to PCR Master Mix
Electrophoresis
The next step was Agarose gel electrophoresis. We can see that there are some of the products, but they do not correspond of all the masses of the products. CfAB and BdhA are both 1000 bp long. 16S is 600 bp long. The reaction have to be repeated.
Agarose electrophoresis: 1-broth, 2-100x diluted broth, 3-DNA, 4-100x diluted DNA
iGEM Project name 1
Main project page
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