Creating devices to test promoters in E. coli strains (devices [http://partsregistry.org/Part:BBa_K177024 BBa_K177024] and [http://partsregistry.org/Part:BBa_K177025 BBa_K177025])

Franek

Tasks:

• Selecting clones with [http://partsregistry.org/Part:BBa_K177024 BBa_K177024] and [http://partsregistry.org/Part:BBa_K177025 BBa_K177025]

Methods:

• Tranformants with [http://partsregistry.org/Part:BBa_K177024 BBa_K177024] and [http://partsregistry.org/Part:BBa_K177025 BBa_K177025] clones were recognised due to the fluorescence under UV light.

• 4 positive [http://partsregistry.org/Part:BBa_K177024 BBa_K177024], and 16 [http://partsregistry.org/Part:BBa_K177025 BBa_K177025] were found.

• Each clone was transferred to new plate with LB, agar-agar and IPTG or 0.2 % arabinose. Liquid cultures were also established.

Results:

• [http://partsregistry.org/Part:BBa_K177024 BBa_K177024] and [http://partsregistry.org/Part:BBa_K177025 BBa_K177025] were successfully created!

Cloning of the mgtc promoter into the pKSII+ plasmid

Kamil

Tasks:

• Bacteria transformation

Methods:

• A 200μl batch of chemocompetent bacteria was transformed with 15μl of ligation mix and incubated overnight on petri dishes containing LB medium supplemented with ampicilin, X-gal and IPTG.

Results:

The transformation yielded only a couple of blue colonies.

Conclusions:

• The transformation needs to be redone, this time using more of the ligation mix.
  

  isolation of pKS/pho

  Kama

  ===
  Assembly of endosomal detection operon
  === '''Marcin''' Task 1: *Isolate plasmids containing the biobricks Methods: *Plasmids weren 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]. *After the isolation 1 ul of each plasmid solution was diluted to 10 ul and loaded into the 1% agarose gel *Electrophoresis condition: voltage - 70V time - 30 min *Next the gel was photographed: [[image:Biobricks_izolacja_15_07_09.png‎|thumb|center|420px|Plasmid samples loaded into the gel]] Legend:
  1-3 - [http://partsregistry.org/Part:BBa_B0032 BBa_B0032] 4-6 - [http://partsregistry.org/Part:BBa_C0040 BBa_C0040] 7-9 - [http://partsregistry.org/Part:BBa_C0051 BBa_C0051] 10-12 - [http://partsregistry.org/Part:BBa_E0032 BBa_E0032] '''Comment''': Isolation of plasmids was successful. ===
  Cloning of p53 coding sequence
  === '''Marcin''' Task: *Cloning p53 coding sequence to pKS plasmid Methods: *Ligation mixture composition: 14 μl digested p53, 1.5 μl digested pKS, 5 μl ligation buffer (Invitrogen), 1 μl ligase T4 * Duration of ligation was about 20 hours; reaction was conducted in 16 °c (approximately). {{WarNotebookEnd}}