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Team:Bielefeld-Germany/Labjournal/week11
From 2012.igem.org
Contents |
Labjournal
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Week 11 (07/09 - 07/15/12)
Monday July 9th
- Ordering primers for isolation of fungal laccases from plasmids, we got from the Ernst-Moritz-Arndt-University in Greifswald. The plasmids contain the cDNA sequences of five different laccases from Trametes versicolor and Pycnoporus cinnabarinus.
- lac5 from Trametes versicolor
- lac10 from Trametes versicolor
- lac13 from Trametes versicolor
- lac20 from Trametes versicolor
- lac35 from Pycnoporus cinnabarinus
- The first primerpairs were designed with standard prefix and suffix sequence and 20 bases complementary to the start and end of the ORF sequences.
- Additional primerpairs were designed with AarI restriction site and Kozak consensus sequence before the first 20 bases from the start of the ORF (forward primers). The reverse primers were desigend with the last 20 bases of the lacase genes and a terminator overhaning end and a AarI restriction site.
Problem: We forgot to delete the signal peptid sequences, which are present in the fungal laccases. Solution: Ordering the forward primers again with the first 20 bases after the signal peptides.
