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Team:Bielefeld-Germany/Labjournal/week17
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Restriktion of pSB1C3+CBDclos with EcoRI+AgeI-HF in Buffer 4 + BSA | Restriktion of pSB1C3+CBDclos with EcoRI+AgeI-HF in Buffer 4 + BSA | ||
To test the Restriktion Enzymes - works | To test the Restriktion Enzymes - works | ||
| + | * '''Team Activity Tests:''' I know, all of you are expecting us to report about some laccase activity now, but today is going to be different. Today we started with a special task: a q-PCR. As written in one of our last labjournal entries we have discussed a lot and searched for reasons why our laccases are not active. We decided that one possible step to find the mistake is to analyze the transcript. The level of mRNA will show us if our plasmid is expressed at all or if there might be something wrong with it. Team Cultivation cultivated small samples of E.coli KRX with plasmids of laccases from the following organisms:''Escherichia coli'', ''Bacillus pumilus'', ''Bacillus halodurans C-125'', ''Xanthomonas campestris pv. campestris B100'' and ''Thermus thermophilus''. As controls a sample with a ligase a plasmid (to ensure the induction works) and ''E. coli KRX'' (without any plasmid) were used. So our first step today was freezing some cell pellets for a RNA isolation and ordering primers. We will go on with our new mission as soon as the primers arrive. | ||
===Wednesday August 22nd=== | ===Wednesday August 22nd=== | ||
Revision as of 14:25, 27 August 2012
Contents |
Labjournal
Week1 Week2 Week3 Week4 Week5 Week6 Week7 Week8 Week9 Week10 Week11 Week12 Week13 Week14 Week15 Week16 Week17 Week18
Week 17 (08/20 - 08/26/12)
Monday August 20th
- Team Site Directed Mutagenesis:
Plasmidisolation of X.camp_2247 1-4 and X.camp_3633 1-4 Testrestriktion with PstI: - all negativ Colony-PCR of T10_NotI (24 Colonies) -all negativ
- Team Cellulose Binding Domain:
CBD: Colony-PCR of 24 Assembly-colonies - all colonies only have CBDclos. Assembly didn't work at all...
Tuesday August 21st
Restriktion of pSB1C3+CBDclos with EcoRI+AgeI-HF in Buffer 4 + BSA To test the Restriktion Enzymes - works
- Team Activity Tests: I know, all of you are expecting us to report about some laccase activity now, but today is going to be different. Today we started with a special task: a q-PCR. As written in one of our last labjournal entries we have discussed a lot and searched for reasons why our laccases are not active. We decided that one possible step to find the mistake is to analyze the transcript. The level of mRNA will show us if our plasmid is expressed at all or if there might be something wrong with it. Team Cultivation cultivated small samples of E.coli KRX with plasmids of laccases from the following organisms:Escherichia coli, Bacillus pumilus, Bacillus halodurans C-125, Xanthomonas campestris pv. campestris B100 and Thermus thermophilus. As controls a sample with a ligase a plasmid (to ensure the induction works) and E. coli KRX (without any plasmid) were used. So our first step today was freezing some cell pellets for a RNA isolation and ordering primers. We will go on with our new mission as soon as the primers arrive.
Wednesday August 22nd
Street Science: Went to Dr. Joe Max Risse today and asked him about the mircoorganisms of the Fermentationgroup. He recommended Euglena gracilis, since it is without risk, big, colorful, and very healthy under the microscop. I asked for the bacteria they have and he told me I could check if their Kocuria rosea is a wild type. In the DMSZ catalog there are three strains of Kocuria rosea and all have been assessed to be of riskgroup 1, which is the safest group of bacteria and means it is unlikely that it will infect humans. I also asked for Penicillium chrysogenum but the one they use is a GVO.
- Team Site Directed Mutagenesis:
Ordered new primers for Xantomonas Campestris SDM, since there is still no positiv coloniy and even gradient-PCR gave the wrong product
- Team Cellulose Binding Domain:
Test-restriktion of all isolated CBDcex- and the GFP_Freiburg-plasmids with NotI - no positiv result Started new and made a gradient PCR for GFP-Freiburg and CBDcex (50-70°C)
