Team:Warsaw/Calendar-Main/1 August 2008

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<p>1. <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#chemotransform">Transformation</a> of E. coli TOP10 strain with ligations (pACYC177+OmpA_alpha + deltaA and pACYC177+OmpA_omega + deltaA). <br>
 
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2. Transformants plating on LB + kanamycin. </p>
 
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<h3> Cloning of truncated fragment of protein A (&Delta;A)</h3>
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Oczyszczanie białek: A-alfa, Z-alfa i Z-omega
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<h4>Michał K.</h4>
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Piotr
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<li> Colony <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> on <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-deltaA-alpha>pACYC177+OmpA_alpha + &Delta;A</a> and <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BOmpA-deltaA-omega>pACYC177+OmpA_omega + &Delta;A</a> colonies of tranformants.</li>
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Zwirowanie bakterii z 1l, sonikacja ich (protokół)
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<li> Gel electrophoresis of PCR products (<a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/1_August_2008#fig1">Fig. 1.</a>).  </li>
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zwirowanie sonikatu
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w przypadku z-alfa i z-omega odżucenie supernatantu, zawieszenie osadu w wodzi i ponowne zwirowanie i odżucenie wody i zawieszenie w wodzie
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w przypadku a-alfa zatrzymanie supernatantu odżucenie osadu
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<br>
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<a name="fig1"><img src="https://static.igem.org/mediawiki/2008/7/7c/Cos.jpg" width=300/></a> <var><b>Fig. 1.</b> Colony PCR to find proper clones of pACYC177_OmpA_alpha+&Delta;A and pACYC177_OmpA_omega+&Delta;A<br>
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Upper gel:<br>
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1. Marker<br>
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2-13. colony PCR products to find proper clone of pACYC177_OmpA_alpha+&Delta;A <br>
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Lower gel:<br>
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1. Marker<br>
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2-11. colony PCR products to find proper clone of pACYC177_OmpA_omega+&Delta;A<br>
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12. positive control - pACYC_OmpA_A_alpha<br>
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13. negative control - pACYC_OmpA_Z_alpha<br></var>

Latest revision as of 18:42, 26 October 2008

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Cloning of truncated fragment of protein A (ΔA)

Michał K.

  1. Colony PCR on pACYC177+OmpA_alpha + ΔA and pACYC177+OmpA_omega + ΔA colonies of tranformants.
  2. Gel electrophoresis of PCR products (Fig. 1.).

Fig. 1. Colony PCR to find proper clones of pACYC177_OmpA_alpha+ΔA and pACYC177_OmpA_omega+ΔA
Upper gel:
1. Marker
2-13. colony PCR products to find proper clone of pACYC177_OmpA_alpha+ΔA
Lower gel:
1. Marker
2-11. colony PCR products to find proper clone of pACYC177_OmpA_omega+ΔA
12. positive control - pACYC_OmpA_A_alpha
13. negative control - pACYC_OmpA_Z_alpha



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