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Team:Bielefeld-Germany/Labjournal/week21
From 2012.igem.org
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===Tuesday September 18th=== | ===Tuesday September 18th=== | ||
===Wednesday September 19th=== | ===Wednesday September 19th=== | ||
| - | * '''Team Cellulose Binding Domain:'''Stopped over-night-''Dpn''I-digestion of S3N10-clos-GFP_his PCR-product and cleaned up in gel. Ligated the PCR-product with itself via Blunt-end-ligation and transformed it into KRX. Plated on Select-Agar. | + | * '''Team Cellulose Binding Domain:''' |
| + | **Stopped over-night-''Dpn''I-digestion of S3N10-clos-GFP_his PCR-product and cleaned up in gel. Ligated the PCR-product with itself via Blunt-end-ligation and transformed it into KRX. Plated on Select-Agar. | ||
**Gradient-PCRs of the CDBcex(T7)+GFP-plasmid and a CBDclos(T7)+GFP-plasmid (unsequenced) with the CBDcex_Freiburg and GFP_Freiburg-compl and the CBDclos_Freiburg and GFP_Freiburg_compl primers, respectively. The CBDcex-setup showed no correct product, but the CBDclos-setup had the right bands at temperatures from 57°C to 64°C; merged the correct fractions and cleaned them up through the gel. | **Gradient-PCRs of the CDBcex(T7)+GFP-plasmid and a CBDclos(T7)+GFP-plasmid (unsequenced) with the CBDcex_Freiburg and GFP_Freiburg-compl and the CBDclos_Freiburg and GFP_Freiburg_compl primers, respectively. The CBDcex-setup showed no correct product, but the CBDclos-setup had the right bands at temperatures from 57°C to 64°C; merged the correct fractions and cleaned them up through the gel. | ||
**Digested the cleaned up product with ''Spe''I and ''Xba''I and ''Dpn''I. | **Digested the cleaned up product with ''Spe''I and ''Xba''I and ''Dpn''I. | ||
| + | **Ligated J61101 and CBDclos with GFP_Freiburg and transformed it into KRX. | ||
===Thursday September 20th=== | ===Thursday September 20th=== | ||
Revision as of 20:54, 19 September 2012
Contents |
Week 21 (09/17 - 09/23/12)
Monday September 17th
Tuesday September 18th
Wednesday September 19th
- Team Cellulose Binding Domain:
- Stopped over-night-DpnI-digestion of S3N10-clos-GFP_his PCR-product and cleaned up in gel. Ligated the PCR-product with itself via Blunt-end-ligation and transformed it into KRX. Plated on Select-Agar.
- Gradient-PCRs of the CDBcex(T7)+GFP-plasmid and a CBDclos(T7)+GFP-plasmid (unsequenced) with the CBDcex_Freiburg and GFP_Freiburg-compl and the CBDclos_Freiburg and GFP_Freiburg_compl primers, respectively. The CBDcex-setup showed no correct product, but the CBDclos-setup had the right bands at temperatures from 57°C to 64°C; merged the correct fractions and cleaned them up through the gel.
- Digested the cleaned up product with SpeI and XbaI and DpnI.
- Ligated J61101 and CBDclos with GFP_Freiburg and transformed it into KRX.
Thursday September 20th
Friday September 21st
Saturday September 22nd
Sunday September 23rd
Sunday
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