Team:Chiba/Calendar-Home/23 October 2008

From 2008.igem.org

(Difference between revisions)
(→Team:Demo-Rs)
Line 4: Line 4:
===Team:Demo-Rs===
===Team:Demo-Rs===
-
<BR>朝
+
===Varying bacterial numbers===
-
<BR>
+
#Receiver(T9002) pre-incubation
-
sender(T9002) グリストをつついて10ml培養×3(LB-Amp)(①washなし、②washあり、③(100:1,1000:1)
+
##Receiver:[http://partsregistry.org/Part:BBa_T9002 BBa_T9002](JW1908)wascultured in 2mL LB-Amp (37&deg;C,12h)
-
<BR>
+
##Pre-incubated Receiver([http://partsregistry.org/Part:BBa_T9002 BBa_T9002](JW1908))was plated so as to produce about 1000 colonies.
-
receiver プレ培したものをコロニーが1000位になるようにプレートに撒く。
+
#Sender(S03623) pre-incubation
-
 
+
##Sender:[http://partsregistry.org/Part:BBa_S03623 BBa_S03623](JW1908) was cultured in 50mL  entrifuge tubes in 10mL of LB-Amp (37&deg;C,12h)(2 tubes)
-
 
+
#Sender Wash
-
夜
+
##Centrifuged 2 tubes containing([http://partsregistry.org/Part:BBa_T9002 BBa_T9002](JW1908))at 20&deg;C,3600rpm for 6min and discarded supernatant.
-
　センダーが濁ったら②③はwash×3
+
##Added 10mL LB-Amp to each tube.
-
　　①は菌液10mlにLB-agar10mlを加える。（Ampも）
+
##Repeated wash twice.
-
　　②はwash後菌液10mlにLB-agarを10ml加える。（Ampも）
+
#Creating bacterial plates
-
　　③(1:100)はwashした菌液を0.1mlとLB-Ampを9.9mlまぜ、そこにLB-agarを10ml加える。
+
##LB-Amp pre-cultured Sender([http://partsregistry.org/Part:BBa_S03623 BBa_S03623](JW1908)) tube 1 (10mL) was mixed with LB-Amp-agar(50&deg;C)(10ml)to produce sender containing bacterialplate-1.
-
　　　(1:1000)はwashした菌液を0.01mlとLB-Ampを9.99mlまぜ、そこにLB-agarを10ml加える。
+
##LB-Amp pre-cultured Sender([http://partsregistry.org/Part:BBa_S03623 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([http://partsregistry.org/Part:BBa_S03623 BBa_S03623](JW1908))containing bacterial plate-2.
-
　　また、ネガコンようとしてLB10mlと、LB-agar10mlを混ぜてプレートにしたものを用意する。
+
##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([http://partsregistry.org/Part:BBa_S03623 BBa_S03623](JW1908) containing bacterial plate-3
-
 
+
#Lifted with nitrocellulose
-
①②③、ネガコンをプレートに移して固まるまで待つ。固まったらreceiver(T9002)をプレートからニトロセルロースで写し取りそれぞれのプレートに2枚ずつはり、その時をスタートとして37℃の培養機の中にいれ、30分ごとにUVを当ててGFPが出ているかどうか確認する。
+
##Receiver([http://partsregistry.org/Part:BBa_T9002 BBa_T9002](JW1908))colony was transfered to a nitrocellulose filter and placed on each of Sender([http://partsregistry.org/Part:BBa_S03623 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).
 +
#Method to detect fluorescence
 +
##Plates cultured at 37&deg;C were exposed to UV (312nm) light once every 30 minutes to observe GFP fluorescence.

Revision as of 21:15, 29 October 2008

>Home | Notebook

22 October 2008 <|> 24 October 2008

Team:Demo-Rs

Varying bacterial numbers

  1. Receiver(T9002) pre-incubation
    1. Receiver:[http://partsregistry.org/Part:BBa_T9002 BBa_T9002](JW1908)wascultured in 2mL LB-Amp (37°C,12h)
    2. Pre-incubated Receiver([http://partsregistry.org/Part:BBa_T9002 BBa_T9002](JW1908))was plated so as to produce about 1000 colonies.
  2. Sender(S03623) pre-incubation
    1. Sender:[http://partsregistry.org/Part:BBa_S03623 BBa_S03623](JW1908) was cultured in 50mL entrifuge tubes in 10mL of LB-Amp (37°C,12h)(2 tubes)
  3. Sender Wash
    1. Centrifuged 2 tubes containing([http://partsregistry.org/Part:BBa_T9002 BBa_T9002](JW1908))at 20°C,3600rpm for 6min and discarded supernatant.
    2. Added 10mL LB-Amp to each tube.
    3. Repeated wash twice.
  4. Creating bacterial plates
    1. LB-Amp pre-cultured Sender([http://partsregistry.org/Part:BBa_S03623 BBa_S03623](JW1908)) tube 1 (10mL) was mixed with LB-Amp-agar(50°C)(10ml)to produce sender containing bacterialplate-1.
    2. LB-Amp pre-cultured Sender([http://partsregistry.org/Part:BBa_S03623 BBa_S03623](JW1908)) tube 2(100μl)was mixed with LB-Amp(9.9ml) and diluted 100-fold. 10ml of this solution was mixed with LB-Amp-agar(50°C)(10ml) and created Sender([http://partsregistry.org/Part:BBa_S03623 BBa_S03623](JW1908))containing bacterial plate-2.
    3. LB-Amp pre-cultured Sender solution-2(10μl) and LB-Amp(9.99ml)was mixed to dilute 1000-fold.10ml of this solution and LB-Amp-agar(50°C)(10ml) was mixed to create Sender([http://partsregistry.org/Part:BBa_S03623 BBa_S03623](JW1908) containing bacterial plate-3
  5. Lifted with nitrocellulose
    1. Receiver([http://partsregistry.org/Part:BBa_T9002 BBa_T9002](JW1908))colony was transfered to a nitrocellulose filter and placed on each of Sender([http://partsregistry.org/Part:BBa_S03623 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).
  6. Method to detect fluorescence
    1. Plates cultured at 37°C were exposed to UV (312nm) light once every 30 minutes to observe GFP fluorescence.
Retrieved from "http://2008.igem.org/Team:Chiba/Calendar-Home/23_October_2008"