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Reporter: Week 6 June 20-25
From 2011.igem.org
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| - | === | + | ===Mutagenesis of XylE, Take 5 Day 5=== |
| + | No growth was seen in any of the three overnight cultures. The most probable reason for the lack of growth in the cultures is the entire colony was used for colony PCR on 6/17, leaving no cells to grow in culture. In order to grow more cells in culture, the three PCR reactions were transformed into competent Escherichia coli cells and plated on chloramphenicol resistant plates. | ||
| + | |||
| + | ===Insert Small Linker into Imp Linker + K3 Vector, Take 2 Day 1=== | ||
| + | The small linker and the Imp Linker + K3 construct were digested with XbaI and AgeI in buffer 4. The digests were ligated together, and the ligation was transformed into Escherichia coli cells and plated onto kanamycin resistant plates. | ||
| + | |||
| + | ===Insert 10 AA Linker into Imp Linker + K3 Vector, Take 3 Day 1=== | ||
| + | The 10 AA linker and the Imp Linker + K3 construct were digested with XbaI and AgeI in buffer 4. The digests were ligated together, and the ligation was transformed into Escherichia coli cells and plated onto kanamycin resistant plates. | ||
| + | |||
| + | ===Insert tev Protease into K3 Vector, Take 4 Day 1=== | ||
| + | No colonies grew from the 6/17 assembly of tev protease and the K3 vector, so the assembly was performed again. The purified tev protease PCR product from 6/4 and the K3 miniprep from 6/13 were digested with XbaI and PstI in buffer 3. The digests were ligated together, and the ligation was transformed into Escherichia coli cells and plated on kanamycin resistant plates. | ||
| + | |||
| + | |||
[[Team:Penn_State/Notebook| Back to Notebook]] | [[Team:Penn_State/Notebook| Back to Notebook]] | ||
Revision as of 17:25, 26 June 2011
Contents |
Monday
Mutagenesis of XylE, Take 5 Day 5
No growth was seen in any of the three overnight cultures. The most probable reason for the lack of growth in the cultures is the entire colony was used for colony PCR on 6/17, leaving no cells to grow in culture. In order to grow more cells in culture, the three PCR reactions were transformed into competent Escherichia coli cells and plated on chloramphenicol resistant plates.
Insert Small Linker into Imp Linker + K3 Vector, Take 2 Day 1
The small linker and the Imp Linker + K3 construct were digested with XbaI and AgeI in buffer 4. The digests were ligated together, and the ligation was transformed into Escherichia coli cells and plated onto kanamycin resistant plates.
Insert 10 AA Linker into Imp Linker + K3 Vector, Take 3 Day 1
The 10 AA linker and the Imp Linker + K3 construct were digested with XbaI and AgeI in buffer 4. The digests were ligated together, and the ligation was transformed into Escherichia coli cells and plated onto kanamycin resistant plates.
Insert tev Protease into K3 Vector, Take 4 Day 1
No colonies grew from the 6/17 assembly of tev protease and the K3 vector, so the assembly was performed again. The purified tev protease PCR product from 6/4 and the K3 miniprep from 6/13 were digested with XbaI and PstI in buffer 3. The digests were ligated together, and the ligation was transformed into Escherichia coli cells and plated on kanamycin resistant plates.
