Team:Chiba/Calendar-Home/21 October 2008

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20 October 2008 <|> 22 October 2008

Varying bacterial numbers
20 October~
 * 1) Pre-incubated Receiver(BBa_T9002(JW1908))was plated so as to produce about 1000 colonies.
 * 2) Sender(S03623) pre-incubation
 * 3) Sender:BBa_S03623(JW1908) was cultured in 50mL entrifuge tubes in 10mL of LB-Amp (37&deg;C,12h)(2 tubes)
 * 4) Sender Wash
 * 5) Centrifuged 2 tubes containing(BBa_T9002(JW1908))at 20&deg;C,3600rpm for 6min and discarded supernatant.
 * 6) Added 10mL LB-Amp to each tube.
 * 7) Repeated wash twice.
 * 8) Creating bacterial plates
 * 9) LB-Amp pre-cultured Sender(BBa_S03623(JW1908)) tube 1 (10mL) was mixed with LB-Amp-agar(50&deg;C)(10ml)to produce sender containing bacterialplate-1.
 * 10) LB-Amp pre-cultured Sender(BBa_S03623(JW1908)) tube 2(100&mu;l)was mixed with LB-Amp(9.9ml) and diluted 100-fold. 10ml of this solution was mixed with LB-Amp-agar(50&deg;C)(10ml) and created Sender(BBa_S03623(JW1908))containing bacterial plate-2.
 * 11) LB-Amp pre-cultured Sender solution-2(10&mu;l) and LB-Amp(9.99ml)was mixed to dilute 1000-fold.10ml of this solution and LB-Amp-agar(50&deg;C)(10ml) was mixed to create Sender(BBa_S03623(JW1908) containing bacterial plate-3
 * 12) Lifted with nitrocellulose
 * 13) Receiver(BBa_T9002(JW1908))colony was transfered to a nitrocellulose filter and placed on each of Sender(BBa_S03623(JW1908))containing bacterial plate (1~3) and Sender-absent negative control plate (t=0). Determined the time required for the colonies to fluoresce depending on the bacterial concentration (100 and 1000-fold dilution).
 * 14) Method to detect fluorescence
 * 15) Plates cultured at 37&deg;C were exposed to UV (312nm) light once every 30 minutes to observe GFP fluorescence.