Template:Team:Bielefeld-Germany/Notebook/Week02

1) Experiment 1
We did it!

2) Experiment 2
Succeeded!

1) Starting with experiment Glycerol dehydrogenase
- Preparing overnight culture ''E. coli'' KRX

2) Experiment 2 - Incubation
Description of experiment 2

1) Glycerol dehydrogenase
- Using overnight culture ''E. coli'' KRX
- Genome Isolation ''E. coli'' KRX using innuPrep Plasmid MiniKit ''JenaAnalytik''
- Using ''E. coli'' KRX Genome DNA for PCR
- PCR-program:
{|cellspacing="0" border="1" |'''Component''' |'''Volume''' |- |H2O |to 50 μL |- |5 x Phusion CC Buffer |10 μL |- |10 mM dNTPs |2 μL |- |Fwd and Rev Primer Mix 10 μM |2,5 μL |- |Tempalte 100 ng |1 μL |- |DMSO |1,5 μL |- |Phusion DNA Polymerase |0,5 μL |} Table PCR-program
- PCR products are used for agarose gel electrophoresis
Table gel-program
- Agarose gel electrophoresis failed, no bands found


2) Experiment 2

1) Glycerol dehydrogenase
- Using'' E. coli'' KRX Genome DNA for PCR
- PCR-program:
Table PCR-components
Table PCR-program
- PCR products are used for agarose gel electrophoresis
Table gel-program
- Agarose gel electrophoresis worked, see image.
Picture Agarosegel glda_1
- Bands are at expected size of 1050 bp


2) Experiment 2

1) Glycerol dehydrogenase
- Repeating agarose gel electrophoresis form 23.05.2013 with more sample volume
- Isolation of bands at 1050 bp using innuPrep Gel Extraction Kit ''JenaAnalytik''
- Measurement of DNA-concentration using nanodrop