Assembly of endosome detection module:

Marcin

Task 1: Transformation of TOP10 bacterial strain with BBa_K177044 on pSB1A3 plasmid Methods:

• Ligation mixture was thermally inactivated (done by Kuba)
• Detailed protocol of transformation is described here

Assembly of fusion proteins

Marcin

Task 1: Isolation of pSB1A3 plasmid containing BBa_K177028 Methods:

• Plasmids were isolated using the A&A plasmid mini kit. Detailed procedure of the isolation is described here

Task 2: Restriction digest of previously isolated plasmids:

Methods:

• Reaction mixture composition:

3 μl purified plasmid DNA product
1 μl EcoRI (Fermentas)
1 μl PstI (Fermentas) 
2 μl Buffer Tango (Fermentas)
13 μl MQ water

• Reactions were carried out 1.5 hour

Results: Really disappointing, none of the attemps to ligate the signal peptide were successful.