Team:Warsaw/Calendar-Main/18 September 2008
From 2008.igem.org
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primers (elongation length 60s) to obtain <a href=http://partsregistry.org/wiki/index.php?title=Part:BBa_K103009>linker_alpha (BBa_K103009)</a> fragment. Reaction was carried out with 25 cycles and in temperature gradient from 55 to 75 °C (four reactions).</li> | primers (elongation length 60s) to obtain <a href=http://partsregistry.org/wiki/index.php?title=Part:BBa_K103009>linker_alpha (BBa_K103009)</a> fragment. Reaction was carried out with 25 cycles and in temperature gradient from 55 to 75 °C (four reactions).</li> | ||
- | <li> Gel electrophoresis of PCR products <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/18_September_2008#fig1">Fig. 1</a>.</li></ol> | + | <li> Gel electrophoresis of PCR products (<a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/18_September_2008#fig1">Fig. 1.</a>).</li></ol> |
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+ | <a name="fig1"><img src="https://static.igem.org/mediawiki/2008/c/c3/Grad2_17_09.jpg" width=300/></a><var><b>Fig. 1. </b>Gradient PCR (temperatures:55-75 °C)<br> | ||
+ | 1. Marker<br> | ||
+ | 2. 55 °C linker_alpha<br> | ||
+ | 3. 60 °C linker_alpha<br> | ||
+ | 4. 65 °C linker_alpha<br> | ||
+ | 5. 70 °C linker_alpha<br></var> | ||
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<h3>Preparation of <a href=http://partsregistry.org/wiki/index.php?title=Part:BBa_K103003>ΔA (BBa_K103003)</a></h3> | <h3>Preparation of <a href=http://partsregistry.org/wiki/index.php?title=Part:BBa_K103003>ΔA (BBa_K103003)</a></h3> | ||
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<a name="fig2"><img src="https://static.igem.org/mediawiki/2008/d/d0/Go_17_09.jpg" width=300/></a><var><b>Fig. 2. PCR to obtain inserts</b><br> | <a name="fig2"><img src="https://static.igem.org/mediawiki/2008/d/d0/Go_17_09.jpg" width=300/></a><var><b>Fig. 2. PCR to obtain inserts</b><br> |
Revision as of 12:51, 27 October 2008
'Hunter and prey' system tests: Competition testsWeronikaInoculation of pACYC177+OmpA_A_alpha, pACYC177+OmpA_Z_omega, pACYC177+OmpA_A_omega and pACYC177+OmpA_Z_alpha + induction using 0.25mM iPTG MutD5 testingPiotrInoculation of mutD5 into 100 ml of LB and rendering them chemocompetent. Electroporation with pACYC177+OmpA_Z_alpha, pACYC177+OmpA_Z_omega, pACYC177+OmpA_A_omega and pACYC177+OmpA_omega_ΔA. Optimisation of primers for preparation of partsMichał K.
1. Marker 2. 55 °C linker_alpha 3. 60 °C linker_alpha 4. 65 °C linker_alpha 5. 70 °C linker_alpha Preparation of ΔA (BBa_K103003)Michał K.
Preparation of linker_alpha (BBa_K103009)Michał K.
Preparation of linker_omega (BBa_K103013)Michał K.
Preparation of OmpA-linker-omega-linker (BBa_K103016)Michał K.
Preparation of OmpA-linker (BBa_K103006)Michał K.
1. Marker 2. deltaA 3 omega 4. ompA_omega Fig. 3. PCR to obtain linker_alpha 1. Marker 2. linker_alpha Fig. 4. Control SacI/NdeI digest of pET15b_OmpA_omega 1. Marker 2. digested pSB1A3 Fig. 5. Control EcoRI/BcuI digest of pSB1A3 1. Marker 2. digested pSB1A3
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