Team:Warsaw/Calendar-Main/4 August 2008

From 2008.igem.org

(Difference between revisions)
Line 20: Line 20:
<ol><li><a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#digest">Digest</a> of pACYC177+OmpA_omega_deltaA_alpha (from 25 July) with BamHI and NotI (BamHI buffer). DNA ends <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#blunting">blunting</a> blunting with Klenow fragment (3 hr). </li>
<ol><li><a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#digest">Digest</a> of pACYC177+OmpA_omega_deltaA_alpha (from 25 July) with BamHI and NotI (BamHI buffer). DNA ends <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#blunting">blunting</a> blunting with Klenow fragment (3 hr). </li>
<li>Gel elctrophoresis and <a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> of proper band - 4300 bp. </li>
<li>Gel elctrophoresis and <a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> of proper band - 4300 bp. </li>
-
<li>Overnight <a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#ligation">ligation</a> of isolated DNA fragment (1 hr). </li>
+
<li>Overnight <a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#ligation">ligation</a> of isolated DNA fragment. </li>
<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#electrotransform">Transformation</a> of E. coli <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> strain with ligation. </li>
<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#electrotransform">Transformation</a> of E. coli <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a> strain with ligation. </li>
<li>Transformants plating on LB + kanamycin. </li></ol>
<li>Transformants plating on LB + kanamycin. </li></ol>

Revision as of 13:11, 12 October 2008

Gallery Bricks Notebook Team Project Home


Previous day
return to main notebook page
Previous entry
next notebook entry

 


Cloning of truncated fragment of protein A

Piotr

Inoculation of some pACYC177+OmpA_alpha + deltaA and pACYC177+OmpA_omega + deltaA colonies of tranformnts.

Checking the expression of omp_omega_A_alpha and omp_A_alpha

Piotr

Inoculation of omp_omega_A_alpha and omp_A_alpha with inductor (0,5 mmol/mL IPTG) and negative control without IPTG.

Checking if omp_omega_A_alpha gives ampicillin resistance

Piotr

Inoculation of omp_omega_A_alpha into various IPTG concentrations: 0, 0.1, 0.25, 0.5, 0.75, 1 mmol/mL.

Preparing pACYC177+OmpA_omega_deltaA construct

Michał K.

  1. Digest of pACYC177+OmpA_omega_deltaA_alpha (from 25 July) with BamHI and NotI (BamHI buffer). DNA ends blunting blunting with Klenow fragment (3 hr).
  2. Gel elctrophoresis and gel-out of proper band - 4300 bp.
  3. Overnight ligation of isolated DNA fragment.
  4. Transformation of E. coli TOP10 strain with ligation.
  5. Transformants plating on LB + kanamycin.