Team:Lethbridge/Notebook

From 2009.igem.org

(Difference between revisions)
(Calendar)
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</head>
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<body>
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<li><a href="https://2009.igem.org/Team:Lethbridge">Home</a></li>
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<li><a href="https://2009.igem.org/Team:Lethbridge/Notebook">Top of Page</a></li>
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== May ==
 +
 +
===May 5===
 +
 +
'''Roxanne, Megan, Alix, Mackenzie, Sebastian'''
 +
 +
*prepared Lb Media in test tubes -prepared 250mL LB agar for plates
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*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===
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'''Roxanne'''
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*Autoclaved LB test tubes, LB agar, Tris-HCl, EDTA, TE, GTE
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*Added ampicillin to LB agar (100ug/mL) -Poured LB + amp plates
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 +
===May 12===
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 +
'''Roxanne'''
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 +
*Prepared LB agar
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*added Tetracyclin (100ug/mL) –
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*Poured LB + ampt plates
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'''
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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
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 +
===May 13===
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'''Roxanne'''
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 +
*Removed test tubes from shaker (16hrs incubation) and placed in deli fridge
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</head>
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<body>
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<ul id="qm0" class="qmmc">
 +
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<li><a href="https://2009.igem.org/Team:Lethbridge">Home</a></li>
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 +
 +
===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
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[[Image:Ohb_PCR-Phusion_(Rox).JPG|250px]]
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<li><a href="https://2009.igem.org/Team:Lethbridge">Home</a></li>
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===May 26===
 +
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'''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
 +
 +
[[Image:Ohb_PCR_(Roxanne).JPG|250px]]
 +
'''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
 +
<html>
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<head>
<html>
<html>

Revision as of 19:21, 21 October 2009


Igembanner.jpg


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

Ohb PCR-Phusion (Rox).JPG

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

Ohb PCR (Roxanne).JPG 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