Team:UIUC Illinois/Notebook

From 2014.igem.org

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Suspended 5 uL of CM34 (antibiotic) + 5 mL of LB + 1 colony of pDCAF3 E. coli<br><br>
Suspended 5 uL of CM34 (antibiotic) + 5 mL of LB + 1 colony of pDCAF3 E. coli<br><br>
Added .5 grams of caffeine + 400 uL of 1M MgSO4 +  20 uL 1M CaCl2 + 40 mL M95x salts +  H2O to YNB +  H2O mixture<br><br>
Added .5 grams of caffeine + 400 uL of 1M MgSO4 +  20 uL 1M CaCl2 + 40 mL M95x salts +  H2O to YNB +  H2O mixture<br><br>
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<ul style="text-index:30px;"><li>Labeled this "M9 media"</li></ul><br>
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<ul style="text-indent:30px;"><li>Labeled this "M9 media"</li></ul><br>
Inoculated frozen stock of E. coli guaB pDCAF3 in 5 mL of M9 media<br><br>
Inoculated frozen stock of E. coli guaB pDCAF3 in 5 mL of M9 media<br><br>
Placed sample in 30 degree celsius shaker for culturing<br><br>
Placed sample in 30 degree celsius shaker for culturing<br><br>
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<h2>Wednesday 11<sup>th</sup> June</h2>
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<a href="https://2014.igem.org/Team:UIUC_Illinois/Project/Overview" target="_blank" class="tbnotelogo PSlogo"> ASDF </a>
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Inoculated CBB1 frozen stock in 5 mL of M9 media<br><br>
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Placed sample in 30 degree celsius shaker<br><br>
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Purified pDCAF plasmid using DNA mini kit<br><br>
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<ul style="text-indent:30px;"><li>Plasmid concentration was 132.9 ng/uL</li></ul><br><br>
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<h2>Wednesday 11<sup>th</sup> June</h2>
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<a href="https://2014.igem.org/Team:UIUC_Illinois/Project/Overview" target="_blank" class="tbnotelogo PSlogo"> ASDF </a>
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Added 10 uL deionized H2O into the J23114 promoter plasmid (Ampᴿ)<br><br>
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Followed bacterial transformation protocol to prepare/grow culture overnight<br><br>
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Inoculated 5 mL of M9 with CBB1 and placed it in 30 degree celsius shaker for 3 days<br><br>
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<ul style="text-indent:30px;"><li>Previous CBB1 culture did not grow, showed slight growth</li><br><br>
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<li>pDCAF3 strain in M9 did grow in the 30 degree celsius shaker</li></ul><br><br>
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Revision as of 01:50, 17 October 2014


Monday 9th June

Made LB broth using 10 grams of bacto agar + 12 grams HiVeg LB broth + 600 mL ddH2O

Autoclaved 1.5 mL Eppendorf tubes as well as LB

Poured plates of LB and chloramphenicol (CAM)

Streaked 2 plates with E. coli DguaB (pDCAF3)

Incubated in 37 degrees celsius

Aliquoted kanamycin (KAN) in four 1mL tubes

Tuesday 10th June

Autoclaved at 1:30pm

  • 0.8 grams of YNB + 156 mL H2O
  • 28 grams of salts + 490 mL H2O

Used sterile filter for 1M MgSO4 and 1M CaCl2

Suspended 5 uL of CM34 (antibiotic) + 5 mL of LB + 1 colony of pDCAF3 E. coli

Added .5 grams of caffeine + 400 uL of 1M MgSO4 + 20 uL 1M CaCl2 + 40 mL M95x salts + H2O to YNB + H2O mixture

  • Labeled this "M9 media"

Inoculated frozen stock of E. coli guaB pDCAF3 in 5 mL of M9 media

Placed sample in 30 degree celsius shaker for culturing

Wednesday 11th June

Inoculated CBB1 frozen stock in 5 mL of M9 media

Placed sample in 30 degree celsius shaker

Purified pDCAF plasmid using DNA mini kit

  • Plasmid concentration was 132.9 ng/uL


Wednesday 11th June

Added 10 uL deionized H2O into the J23114 promoter plasmid (Ampᴿ)

Followed bacterial transformation protocol to prepare/grow culture overnight

Inoculated 5 mL of M9 with CBB1 and placed it in 30 degree celsius shaker for 3 days

  • Previous CBB1 culture did not grow, showed slight growth


  • pDCAF3 strain in M9 did grow in the 30 degree celsius shaker





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