Team:Warsaw/Calendar-Main/4 August 2008

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<h3>Checking <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-omega-A-alpha>OmpA_omega_&Delta;A_alpha</a> expression</h3><h4>Piotr</h4>
<h3>Checking <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-omega-A-alpha>OmpA_omega_&Delta;A_alpha</a> expression</h3><h4>Piotr</h4>
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<p>Inoculation of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-omega-A-alpha>OmpA_omega_&Delta;A_alpha</a> and <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-A-alpha>OmpA_A_alpha</a> with inductor (0,5 mmol/mL IPTG) and negative control without IPTG.</p>
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<p>Inoculation of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-omega-A-alpha>OmpA_omega_&Delta;A_alpha</a> with inductor (0,5 mmol/mL IPTG) and negative control without IPTG.</p>

Latest revision as of 10:50, 29 October 2008

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Cloning of truncated fragment of protein A (ΔA)

Piotr

Inoculation of some pACYC177+OmpA_alpha + ΔA and pACYC177+OmpA_omega + ΔA colonies of tranformants.

Checking OmpA_omega_ΔA_alpha expression

Piotr

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

Checking if OmpA_omega_ΔA_alpha gives ampicillin resistance

Emilia

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

Preparing pACYC177+OmpA_omega_ΔA construct

Michał K.

  1. Digest of pACYC177+OmpA_omega_ΔA_alpha (from 25 July) with BamHI and NotI (BamHI buffer). DNA ends blunting with Klenow fragment (3 hr).
  2. Gel elctrophoresis (Fig. 1) and gel-out of proper band - 4300 bp.
  3. Overnight ligation of isolated DNA fragment.
Fig. 1. Gel-out of pACYC177_OmpA_omega_deltaA_alpha digested with NotI/BamHI after blunting ends.
1. Marker
2. pACYC177_OmpA_omega_deltaA_alpha