Team:Warsaw/Calendar-Main/29 July 2008

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<a name="fig1"><img src="https://static.igem.org/mediawiki/2008/6/6a/Po_go.jpg" width=300/></a> <var><b>Fig. 1. </b>Control SacI/NotI digests of isolated plasmids<br>  
<a name="fig1"><img src="https://static.igem.org/mediawiki/2008/6/6a/Po_go.jpg" width=300/></a> <var><b>Fig. 1. </b>Control SacI/NotI digests of isolated plasmids<br>  
1. Marker<br>
1. Marker<br>
-
2. digested pKS_omega_deltaA<br>  
+
2. digested pKS_omega_&Delta;A<br>  
3. digested pACYC_Omp_alpha<br></var>
3. digested pACYC_Omp_alpha<br></var>

Revision as of 18:59, 26 October 2008

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

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