Team:Groningen/Notebook/Wetwork 2July2012

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Arjan/Renske
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Arjan/Renske<br><br>
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'''Micro-Array cell harvesting'''
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Micro-Array cell harvesting<br><br>
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O/N culture diluted to OD = 0.326 (9:20h). Grown until OD = 0.775 (11:05).  
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O/N culture diluted to OD = 0.326 (9:20h). Grown until OD = 0.775 (11:05). <br>
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3.6 mL brought to flask with magnet stirrer and 45 mL LB, rest put back in 37 degrees shaker for next experiment. OD = 0.095. 11:11 Micro-array harvest setup started in 37 degrees room. Stirrer on 8. Air blown over fresh meat* (kept on ice).
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3.6 mL brought to flask with magnet stirrer and 45 mL LB, rest put back in 37 degrees shaker for next experiment. OD = 0.095. 11:11 Micro-array harvest setup started in 37 degrees room. Stirrer on 8. Air blown over fresh meat* (kept on ice).<br>
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At 13:05 cells harvested by centrifuging (1 min, 10000 rpm, 4 degrees C) and put in -80.
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At 13:05 cells harvested by centrifuging (1 min, 10000 rpm, 4 degrees C) and put in -80.<br>
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12:20: remaining culture (OD = 1,78) diluted to OD = 0.311. 2 hours, 37 degrees (OD = 1,2). Brought to to flask with magnet stirrer and 45 mL LB, OD = 0.118. 13:20 Micro-array harvest setup started in 37 degrees room. Stirrer on 8. Air blown over rotten meat* (kept on ice).
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12:20: remaining culture (OD = 1,78) diluted to OD = 0.311. 2 hours, 37 degrees (OD = 1,2). Brought to to flask with magnet stirrer and 45 mL LB, OD = 0.118. 13:20 Micro-array harvest setup started in 37 degrees room. Stirrer on 8. Air blown over rotten meat* (kept on ice).<br><br>
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*minced meat, 30% beef, 70% pork, bougth at Jumbo grocery store. Rotten meat was kept for 14 days in fridge, fresh meat was bought today.
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*minced meat, 30% beef, 70% pork, bougth at Jumbo grocery store. Rotten meat was kept for 14 days in fridge, fresh meat was bought today.<br><br>
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Emeraldo/Nisa
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Emeraldo/Nisa<br><br>
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1.) Plasmid isolation of BBa_I742123; concentration: 280ng/ul
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1.) Plasmid isolation of BBa_I742123; concentration: 280ng/ul<br>
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2.) Linearization of isolated plasmid with SpeI. Result: correct length
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2.) Linearization of isolated plasmid with SpeI. Result: correct length<br>
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3.) B. subtilis transformation with: BBa_K090403 (amyE integration backbone), BBa_I742123 (multi host vector plasmid), BBa_K274002 (violacein pigment). The yellow pigment and red pigment are not used here, because of the amp resistance gene: B. subtilis can't receive amp resistant gene. (the pigment genes must be inserted to another backbonoe with different antibiotic resistance).
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3.) B. subtilis transformation with: BBa_K090403 (amyE integration backbone), BBa_I742123 (multi host vector plasmid), BBa_K274002 (violacein pigment). The yellow pigment and red pigment are not used here, because of the amp resistance gene: B. subtilis can't receive amp resistant gene. (the pigment genes must be inserted to another backbonoe with different antibiotic resistance).<br>
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<A HREF="https://2012.igem.org/Team:Groningen/Notebook"><FONT COLOR=#ff6700>Back to notebook</FONT></A>
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Revision as of 18:10, 15 September 2012




Arjan/Renske

Micro-Array cell harvesting

O/N culture diluted to OD = 0.326 (9:20h). Grown until OD = 0.775 (11:05).
3.6 mL brought to flask with magnet stirrer and 45 mL LB, rest put back in 37 degrees shaker for next experiment. OD = 0.095. 11:11 Micro-array harvest setup started in 37 degrees room. Stirrer on 8. Air blown over fresh meat* (kept on ice).
At 13:05 cells harvested by centrifuging (1 min, 10000 rpm, 4 degrees C) and put in -80.
12:20: remaining culture (OD = 1,78) diluted to OD = 0.311. 2 hours, 37 degrees (OD = 1,2). Brought to to flask with magnet stirrer and 45 mL LB, OD = 0.118. 13:20 Micro-array harvest setup started in 37 degrees room. Stirrer on 8. Air blown over rotten meat* (kept on ice).

*minced meat, 30% beef, 70% pork, bougth at Jumbo grocery store. Rotten meat was kept for 14 days in fridge, fresh meat was bought today.

Emeraldo/Nisa

1.) Plasmid isolation of BBa_I742123; concentration: 280ng/ul
2.) Linearization of isolated plasmid with SpeI. Result: correct length
3.) B. subtilis transformation with: BBa_K090403 (amyE integration backbone), BBa_I742123 (multi host vector plasmid), BBa_K274002 (violacein pigment). The yellow pigment and red pigment are not used here, because of the amp resistance gene: B. subtilis can't receive amp resistant gene. (the pigment genes must be inserted to another backbonoe with different antibiotic resistance).

Back to notebook