Team:Lethbridge/Notebook

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Calendar

May

May 5

Roxanne, Megan, Alix, Mackenzie, Sebastian

prepared Lb Media in test tubes -prepared 250mL LB agar for plates

Prepared 500nM Tris-HCl (pH 8.0), EDTA (pH 8.0, 500nm) TE Buffer (pH 8.0), GTE (Solution I), 2M NaOH, 10% SDS

May 6

Roxanne

Autoclaved LB test tubes, LB agar, Tris-HCl, EDTA, TE, GTE

Added ampicillin to LB agar (100ug/mL) -Poured LB + amp plates

May 12

Roxanne

Prepared LB agar
added Tetracyclin (100ug/mL) –
Poured LB + ampt plates

Roxanne, Alix, Ashley, Megan, Mackenzie, Kirsten

transferred pE43 (DH5α) cells from gylcerol stock to 5mL LB+Tet test tube (x8)
placed in orbital shaker overnight at 37°C

May 13

Roxanne

Removed test tubes from shaker (16hrs incubation) and placed in deli fridge

May 14

Roxanne, Alix, Ashley, Mackenzie, Kirsten

Preformed alkaline lysis miniprep on pE43 plasmid -lost plasmid from 2 tubes

obtained 6X pE43 plasmid (2X ohbR, 2x ohbB, ohbA, ohbC)

May 19

Roxanne, Alix, Ashley, Kirsten, Megan, Mackenzie

Gel Electrophoresis : 1% agarose in TAE

Lane 1: Ladder

Lane 2: ohbR1

Lane 3: ohbR2

Lane 4: ohbC

Lane 5: ohbB1

Lane 6: ohbB2

Lane 7:ohbA

Lane 8:ohbC

Lane 9: ohbB1

Lane 10:ohbB2

DNA quantification by UV spec 260nm : 2.013 280nm: 1.105 260/280 : 1.984 Concentration: 2517µg/mL PCR of ohbA, (ohbB)x2, ohbC, (ohbR)x2

Took picture of the gel - it did not turn out.

May 21

Roxanne

Made 50X TAE Buffer
Running PCR amplicons & RS/pSB1A2 ligation product on a 1% agarose gel
Jeff gave tutorial on primer design and quick-change mutagenesis to Kirsten, Ashley, Mackenzie, Fan, Lisza, Roxanne

May 26

Roxanne, Alix, Megan, Mackenzie, Kirsten

Gel did not turn out gave tutorial on open wetware and the igem registry

May 28

Roxanne

reran the gel from may 19th, only primer dimers present

Roxanne, Ashley, Alix, Kirsten, Mackenzie, Megan

Redoing the PCR of ohbA, ohbB, ohbC and OhbR with Phusion DNA Polymerase

Roxanne

refilled the tip boxes

May 29

Roxanne

prepared glycerol stocks for autoclaving