Team:Warsaw/Calendar-Main/29 July 2008

From 2008.igem.org

(Difference between revisions)
 
Line 21: Line 21:
<li>Transformants plating on LB + kanamycin.</li></ol>  
<li>Transformants plating on LB + kanamycin.</li></ol>  
</p>
</p>
-
<a name="fig1"><img src="https://static.igem.org/mediawiki/2008/6/6a/Po_go.jpg" width=300/></a> <var><b>Fig. 1. </b> SacI/NotI digests of isolated plasmids<br>  
+
<a name="fig1"><img src="https://static.igem.org/mediawiki/2008/6/6a/Po_go.jpg" width=200/></a> <var><b>Fig. 1. </b> SacI/NotI digests of isolated plasmids<br>  
1. Marker<br>
1. Marker<br>
2. digested pKS_omega_&Delta;A<br>  
2. digested pKS_omega_&Delta;A<br>  

Latest revision as of 16:55, 28 October 2008

Gallery Bricks Notebook Team Project Home


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

 



Purification of proteins: Z-alpha and Z-omega

Piotr

pET15b+Z_alpha and pET15b+Z_omega in Rosetta strain inoculated in liquid LB with chloramphenicol and ampicillin.

Cloning of omega_ΔA DNA fragment to pACYC177+OmpA_alpha

Michał K.

  1. Isolation of pKS+omega_ΔA from culture inoculated on previous day.
  2. Digest of pKS+omega_ΔA with SacI and NotI (BamHI buffer).
  3. Digest (SacI and NotI) and dephosphorylation (CIAP) of pACYC177+OmpA_alpha.
  4. Gel electrophoresis and gel-out of 4300 bp (pACYC177+OmpA_alpha) and 550 bp (omega_ΔA) bands (Fig. 1.).
  5. Ligation of DNA fragments from 2. and 3. (1 hr).
  6. Transformation of E. coli TOP10 strain with ligation.
  7. Transformants plating on LB + kanamycin.

Fig. 1. SacI/NotI digests of isolated plasmids
1. Marker
2. digested pKS_omega_ΔA
3. digested pACYC_Omp_alpha

Retrieved from "http://2008.igem.org/Team:Warsaw/Calendar-Main/29_July_2008"