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

Jarek

Tasks:

• Electrophoretical segregation of PCR products from previous day in 0,8% agarosis gel

• Preparation of another PCR reaction with L.monocytogenes genome DNA

Results:

• I propably took the wrong genome DNA sample for the previous reaction, there are products but none of them has the correct length.

Assembly of endosomal detection operon

Marcin

Task 1:

• Prepare the bacterial cultures for isolation of following constructs:
  • [http://partsregistry.org/Part:pSB1A3pSB1A3] plasmid containing cro coding sequence
  • [http://partsregistry.org/Part:pSB1A3pSB1A3] plasmid containing Bax coding sequence

Task 2:

• Isolate the plasmids containing previously described constructs

Methods: Plasmids were isolated using the A&A plasmid mini kit. Detailed procedure of the isolation is described[http://aabiot.com/products/dna_purification/plasmid_dna/plasmid_mini/protocol_plasmid_mini.pdf here]

Task 3:

• Digest previously isolated plasmids and verify the correctness of the ligation:

Methods:

• Reaction mixture composition:

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

• Digest was performed 90 minutes

Results:

Comments:

• All samples of cro CDS on [http://partsregistry.org/Part:pSB1A3pSB1A3] are only empty plasmids
• Samples of Bax CDS have biobricks but I am unable to determine whether this biobricks is Bax CDS or [http://partsregistry.org/Part:BBa_E0010BBa_E0010] due to contamination with the latter one. It is obligated to prepare another digestion to reveal the identity of the insert