"
Team:Cornell/project/wetlab/chassis
From 2014.igem.org
(Difference between revisions)
(Insert DH5a) |
|||
| Line 10: | Line 10: | ||
<div class="container" id="top"> | <div class="container" id="top"> | ||
<div class="row"> | <div class="row"> | ||
| - | <div class="col-md- | + | <div class="col-md-9 col-xs-18"> |
<h1 style="margin-top: 0px;"><i>E. coli DH5α</i></h1> | <h1 style="margin-top: 0px;"><i>E. coli DH5α</i></h1> | ||
<i>E. coli</i> is a gram-negative, facultative anaerobic, rod-shaped bacterium that is the most widely studied prokaryotic model organism for work with recombinant DNA. We are using electrocompotent and chemically compotent DH5α strains that contain recA1 and endA1 mutations that increase insert stability and improve the quality of plasmid DNA prepared from minipreps <sup>[1]</sup>. | <i>E. coli</i> is a gram-negative, facultative anaerobic, rod-shaped bacterium that is the most widely studied prokaryotic model organism for work with recombinant DNA. We are using electrocompotent and chemically compotent DH5α strains that contain recA1 and endA1 mutations that increase insert stability and improve the quality of plasmid DNA prepared from minipreps <sup>[1]</sup>. | ||
<br><br> | <br><br> | ||
| - | + | <h1><i>E. coli BL21</i></h1> | |
| - | + | We are also using BL21 competent strains of <i>E. coli</i>, which are designed for high-efficiency protein expression of any gene under control of the T7 bacteriophage promoter system. T7 RNA Polymerase is regulated by arabinose induction and glucose inhibition of the <i>ara</i>BAD promoter in its chromosomal DNA. Proteins under control of T7 promoter driven vectors are then expressed at high levels. The cells also contain the T7 lysozyme gene within the pLysS plasmid, which suppresses T7 RNA polymerase activity caused by leaky basal expression <sup>[2]</sup>. | |
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
</div> | </div> | ||
<div class="col-md-3 col-xs-5"> | <div class="col-md-3 col-xs-5"> | ||
<div href="#" class="thumbnail"> | <div href="#" class="thumbnail"> | ||
| - | <img src="https://static.igem.org/mediawiki/ | + | <<img src="https://static.igem.org/mediawiki/2013/6/6e/Bl21_rotate.png" style="padding-top:40px; max-height:none">> |
</div> | </div> | ||
</div> | </div> | ||
Revision as of 02:32, 3 October 2014
Wet Lab
E. coli DH5α
E. coli is a gram-negative, facultative anaerobic, rod-shaped bacterium that is the most widely studied prokaryotic model organism for work with recombinant DNA. We are using electrocompotent and chemically compotent DH5α strains that contain recA1 and endA1 mutations that increase insert stability and improve the quality of plasmid DNA prepared from minipreps [1].E. coli BL21
We are also using BL21 competent strains of E. coli, which are designed for high-efficiency protein expression of any gene under control of the T7 bacteriophage promoter system. T7 RNA Polymerase is regulated by arabinose induction and glucose inhibition of the araBAD promoter in its chromosomal DNA. Proteins under control of T7 promoter driven vectors are then expressed at high levels. The cells also contain the T7 lysozyme gene within the pLysS plasmid, which suppresses T7 RNA polymerase activity caused by leaky basal expression [2].
<
>
>
Far far away, behind the word mountains, far from the countries Vokalia and Consonantia, there live the blind texts. Separated they live in Bookmarksgrove right at the coast of the Semantics, a large language ocean. A small river named Duden flows by their place and supplies it with the necessary regelialia.
But I must explain to you how all this mistaken idea of denouncing pleasure and praising pain was born and I will give you a complete account of the system, and expound the actual teachings of the great explorer of the truth, the master-builder of human happiness.
Header 2
The quick, brown fox jumps over a lazy dog. DJs flock by when MTV ax quiz prog. Junk MTV quiz graced by fox whelps. Bawds jog, flick quartz, vex nymphs. Waltz, bad nymph, for quick jigs vex! Fox nymphs grab quick-jived waltz. Brick quiz whangs jumpy veldt fox. Bright vixens jump; dozy fowl quack.
No one rejects, dislikes, or avoids pleasure itself, because it is pleasure, but because those who do not know how to pursue pleasure rationally encounter consequences that are extremely painful. Nor again is there anyone who loves or pursues or desires to obtain pain of itself, because it is pain, but because occasionally circumstances occur in which toil and pain can procure him some great pleasure. To take a trivial example, which of us ever undertakes laborious physical exercise, except to obtain some advantage from it? But who has any right to find fault with a man who chooses to enjoy a pleasure that has no annoying consequences, or one who avoids a pain that produces no resultant pleasure? On the other hand, we denounce with righteous indignation and dislike men who are so beguiled and demoralized by the charms of pleasure of the moment, so blinded by desire, that they cannot foresee.
Header 3
The quick, brown fox jumps over a lazy dog. DJs flock by when MTV ax quiz prog. Junk MTV quiz graced by fox whelps. Bawds jog, flick quartz, ex nymphs. Waltz, bad nymph, for quick jigs vex! Fox nymphs grab quick-jived waltz. Brick quiz whangs jumpy veldt fox. Bright vixens jump; dozy fowl quack.References
- Ref 1
- Ref 2
- Ref 3
