Team:Aachen/TestBench2

From 2014.igem.org

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{{Team:Aachen/newHeader}}
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{{Team:Aachen/Header}}
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__TOC__
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Wonderful woman. Very free-spirited. We're all very fond of her. And I'm talkin' about the Dude here —sometimes there's a man who, wal, he's the man for his time'n place, he fits right in there—and that's the Dude, in Los Angeles. They call Los Angeles the City of Angels. I didn't find it to be that exactly, but I'll allow as there are some nice folks there. 'Course, I can't say I seen London, and I never been to France, and I ain't never seen no queen in her damn undies as the fella says. But I'll tell you what, after seeing Los Angeles and thisahere story I'm about to unfold —wal, I guess I seen somethin' ever' bit as stupefyin' as ya'd see in any a those other places, and in English too, so I can die with a smile on my face without feelin' like the good Lord gypped me.
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= Media =
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{{Team:Aachen/Gallery
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== LB medium ==
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|file=Aachen_MakerFaire_01.JPG
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# weight components
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|file=Aachen_MakerFaire_02.JPG
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::* '''5 g/L NaCl'''
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}}
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::* '''10 g/L tryptone'''
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::* '''5 g/L yeast extract'''
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::* (15 g/L agar for plates)
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# fill up to 1 L with deionized water
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* '''mix well''' by shaking
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# autoclave
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## autoclaving tape, caps slightly unscrewed
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## base of the pot has to be covered with deionized water
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## close lid
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## heat '''level 3 until the pressure valve opens'''
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## reduce '''heat level to 1.5'''
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## set timer to '''20 minutes'''
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## turn heater off
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## '''wait until the pressure valve retracts''' (30-45 minutes)
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## open, close caps & shake
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# for plates, wait until you can touch the bottle ('''<60 °C''', clean bench!)
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# add antibiotics and '''shake''' (gloves!)
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== Hartmans minimal medium (HM) ==
 
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They won't hurt us, Donny. These men are cowards. Fine, Dude. As if it's impossible to get some nail polish, apply it to someone else's toe. Donny was a good bowler, and a good man. He was… He was one of us. He was a man who loved the outdoors, and bowling, and as a surfer explored the beaches of southern California from Redondo to Calabassos. And he was an avid bowler. And a good friend. He died—he died as so many of his generation, before his time. In your wisdom you took him, Lord. As you took so many bright flowering young men, at Khe San and Lan Doc.
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{| class="wikitable"
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Look, I've got certain information, certain things have come to light, and uh, has it ever occurred to you, man, that given the nature of all this new shit, that, uh, instead of running around blaming me, that this whole thing might just be, not, you know, not just such a simple, but uh—you know? Your goons'll be able to get it off him, mean he's only fifteen and he's flunking social studies. So if you'll just write me a check for my ten per cent… of half a million… fifty grand.
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|-
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! Component !! supplement for 1x HM [g/L] !! final 1x concentration
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|-
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| Casamino acids || 2 || 2 g/L
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|-
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| Thiamine hydrochloride || 0.3 || 300 mg/L
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|-
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| MgSO{{sub|4}}/MgSO{{sub|4}}*7H{{sub|2}}O || 0.242 / 0,494 || 2 mmol/L
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|-
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| CaCl{{sub|2}} || 0.011 || 0.1 mmol/L
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|}
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= Agar Chips =
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Whose toe was it, Walter? Is this yours, Larry? Is this your homework, Larry? Your "revolution" is over, Mr. Lebowski! Condolences! The bums lost! Hello, Pilar? My name is Walter Sobchak, we spoke on the phone, this is my associate Jeffrey Lebowski. Blow on them. Strong men also cry… Strong men also cry. Our basic freedoms. Uh, yeah. Probably a vagrant, slept in the car. Or perhaps just used it as a toilet, and moved on.
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{{Team:Aachen/Figure|Aachen_Team_Logo_01.png|title=Figure 1|subtitle=This is our team logo|width=200px}}
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= Transformation =
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Fuckin' A. No ma'am, I didn't mean to give the impression that we're police exactly. We're hoping that it will not be necessary to call the police. Hey, man, if my fucking ex-wife asked me to take care of her fucking dog while she and her boyfriend went to Honolulu, I'd tell her to go fuck herself. They finally did it. They killed my fucking car.
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== Heat Shock ==
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# thaw cells on ice
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# add 1&nbsp;µl of plasmid DNA
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# incubate on ice for 30'
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# heat shock at 42&nbsp;°C for 60''
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# incubate on ice for 5'
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# add 200&nbsp;µl of SOC media
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# incubate at 37&nbsp;°C for 2&nbsp;h
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# plate 20&nbsp; and 200&nbsp;µl on plates with the appropiate antibiotic
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== Electroporation ==
 
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= PCR =
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{{Team:Aachen/Footer}}
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== Colony PCR ==
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== QuikChange ==
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= Clonings =
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== Restriction Digest ==
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== Ligation ==
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== Gibson Assembly ==
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{{Team:Aachen/newFooter}}
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Revision as of 20:46, 9 October 2014

Wonderful woman. Very free-spirited. We're all very fond of her. And I'm talkin' about the Dude here —sometimes there's a man who, wal, he's the man for his time'n place, he fits right in there—and that's the Dude, in Los Angeles. They call Los Angeles the City of Angels. I didn't find it to be that exactly, but I'll allow as there are some nice folks there. 'Course, I can't say I seen London, and I never been to France, and I ain't never seen no queen in her damn undies as the fella says. But I'll tell you what, after seeing Los Angeles and thisahere story I'm about to unfold —wal, I guess I seen somethin' ever' bit as stupefyin' as ya'd see in any a those other places, and in English too, so I can die with a smile on my face without feelin' like the good Lord gypped me.


They won't hurt us, Donny. These men are cowards. Fine, Dude. As if it's impossible to get some nail polish, apply it to someone else's toe. Donny was a good bowler, and a good man. He was… He was one of us. He was a man who loved the outdoors, and bowling, and as a surfer explored the beaches of southern California from Redondo to Calabassos. And he was an avid bowler. And a good friend. He died—he died as so many of his generation, before his time. In your wisdom you took him, Lord. As you took so many bright flowering young men, at Khe San and Lan Doc.

Look, I've got certain information, certain things have come to light, and uh, has it ever occurred to you, man, that given the nature of all this new shit, that, uh, instead of running around blaming me, that this whole thing might just be, not, you know, not just such a simple, but uh—you know? Your goons'll be able to get it off him, mean he's only fifteen and he's flunking social studies. So if you'll just write me a check for my ten per cent… of half a million… fifty grand.

Whose toe was it, Walter? Is this yours, Larry? Is this your homework, Larry? Your "revolution" is over, Mr. Lebowski! Condolences! The bums lost! Hello, Pilar? My name is Walter Sobchak, we spoke on the phone, this is my associate Jeffrey Lebowski. Blow on them. Strong men also cry… Strong men also cry. Our basic freedoms. Uh, yeah. Probably a vagrant, slept in the car. Or perhaps just used it as a toilet, and moved on.

Fuckin' A. No ma'am, I didn't mean to give the impression that we're police exactly. We're hoping that it will not be necessary to call the police. Hey, man, if my fucking ex-wife asked me to take care of her fucking dog while she and her boyfriend went to Honolulu, I'd tell her to go fuck herself. They finally did it. They killed my fucking car.


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