Assembly of endosomal detection operon

Marcin

Task 1:

• Restriction digest of followed costruct: BBa_K177037 from the pSB1A3 plasmid

Methods:

• Reaction mixture composition:

 
1. μl purified plasmid DNA product
1 μl XbaI (Fermentas)
1 μl PstI (Fermentas) 
2 μl Buffer Tango (Fermentas)
15 μl MQ water

• Digest was performed about six hours and subsequently thermal inactivated

Results

• All isolated plasmids have no insert

Task 2:

• Prepare the bacterial cultures for isolation of on pSB1A3 plasmid containing BBa_K177035

Acquiring the internaline A gene from Listeria monocytogenes st. EDG-e genome

Jarek

Tasks:

• Preparation of PCR reaction with L.monocytogenes st. EGD-e genome DNA
• Separation of PCR products on the 0,8% agarose gel
• Acquiring the pure DNA sample from PCR reaction with Clean-Up kit

Cloning of the mgtc promoter into the pSB1A3 plasmid

Kamil

Tasks:

• Colonies transfer
• Liquid cultures establishment

Methods:

• Appropriate colonies (both of them) were transferred to a new plate containing ampicilin
• Liquid cultures were established in 5 ml LB medium supplemented with ampicilin and incubated overnight with areation

Cloning of the cro-box into the pSB1A3 plasmid

Kamil

Tasks:

• Colonies transfer
• Liquid cultures establishment

Methods:

• Appropriate colonies (all 7 of them) were transferred to a new plate containing ampicilin
• Liquid cultures were established in 5 ml LB medium supplemented with ampicilin and incubated overnight with areation

Cloning switch 1 regulatory parts [ K177012-PcI.RBS.LacI, K177033-PcI.RBS.LacI.PcI.RBS.RFP.terminator, K177011-PLacI.RBS.cI.terminator, K177038-PLacI.RBS.cI.terminator.PLacI.RBS.GFP.terminator ] into two compatible low copy number plasmids of different antibiotic resistance

Ania

Tasks: