lab 208

Main purpose

Make PCR on Nir with genomic DNA template. Amplify the pZE21 backbone with USER-primers for construction of the arabinose inducible system. Put His-tag on the cycAX construct.

Who was in the lab

Gosia, Kristian, Henrike, Julia

Procedure

PCR

• pZE21::GFP SF and pZE21::RFP in duplicates with primers pair 13 on 55C and extension time 3:00.
• cycAX in triplicates with primer pair 33 2 samples on 62C and 3:30 extension and 1 sample on 55C and extension 3:00.
• None negative was made for each containing same things as real samples just MQ instead of template DNA.

Results

Gel on PCR from yesterday

• 1kb ladder
• Nir 1uL genomic template
• Nir 3uL genomic template
• Nir 7uL....
• Nir 10uL....
• Nir 12uL....
• Nir 15uL....
• Nir 1uL HindIII treated genomic template
• Nir 3uL HindIII treated genomic template
• Nir 7uL....
• Nir 10uL....
• Nir 12uL....
• Nir 15uL....
• Negative control
• 1kb ladder

Conclusion

No right bands where acquired from PCR-amplification with genomic DNA template.

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