Team:Warsaw/Calendar-Main/1 August 2008

From 2008.igem.org

(Difference between revisions)
 
(21 intermediate revisions not shown)
Line 4: Line 4:
<html>
<html>
-
<p><ol>
 
-
<li> <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#chemotransform">Transformation</a> of <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top">TOP10</a> strain with ligations (pACYC177+OmpA_alpha + deltaA and pACYC177+OmpA_omega + deltaA). </li>
 
-
<li> Transformants plating on LB + kanamycin. </li></ol></p>
 
 +
<h3> Cloning of truncated fragment of protein A (&Delta;A)</h3>
 +
<h4>Michał K.</h4>
 +
<ol>
 +
<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>
 +
<li> Gel electrophoresis of PCR products (<a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/1_August_2008#fig1">Fig. 1.</a>).  </li>
 +
</ol>
-
Sprawdzenie czy degradacja fuzji z Ompa jest wynikiem działania proteaz lon iompt (obecne w top10)
+
<br>
-
PIOTR
+
<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>
-
 
+
Upper gel:<br>
-
Zwirowanie, zawieszenie, dodanie buforu lizującego (skład), zagotowanie, nałożenie na żel poliakryl, transfer na nitrocelulozę, blokowanie, przeciwciała anty-A, płukanie, przeciwciała anty-królik, wywołanie BCIP i NBT (omp_omega_A_alfa, z i bez indukcji, omp_A_alfa)
+
1. Marker<br>
-
 
+
2-13. colony PCR products to find proper clone of pACYC177_OmpA_alpha+&Delta;A <br>
-
(Zdjęcie żelu)
+
Lower gel:<br>
-
 
+
1. Marker<br>
-
Brak rużnic ekspresji i degradacji w top10 i rosettach => degradacja nie wynika z działania proteaz- z czego?
+
2-11. colony PCR products to find proper clone of pACYC177_OmpA_omega+&Delta;A<br>
-
 
+
12. positive control - pACYC_OmpA_A_alpha<br>
 +
13. negative control - pACYC_OmpA_Z_alpha<br></var>

Latest revision as of 18:42, 26 October 2008

Gallery Bricks Notebook Team Project Home


Previous day
return to main notebook page
Previous entry
Interactive list of topics
next notebook entry

 



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



Retrieved from "http://2008.igem.org/Team:Warsaw/Calendar-Main/1_August_2008"