IGEM:University of East Anglia (UEA), Norwich, UK/2009/Notebook/NRP-UEA-Norwich iGEM/2014/07/10: Difference between revisions
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|style="background-color: #EEE"|[[Image:igem-logo-150px.png|150px]]<span style="font-size:22px;"> iGEM Project name 1</span> | |style="background-color: #EEE"|[[Image:igem-logo-150px.png|150px]]<span style="font-size:22px;"> iGEM Project name 1</span> | ||
|style="background-color: #F2F2F2" align="center"| | |style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} | ||
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7. Store plants in controlled environment at 19-23 °C for 3 days. | 7. Store plants in controlled environment at 19-23 °C for 3 days. | ||
-Ligation reaction to join PSB1C3 and GG compatible ends | |||
•PCR program? | |||
•Transfored into E.coli | |||
•Spread plated onto CAM plates | |||
Latest revision as of 00:06, 27 September 2017
 iGEM Project name 1
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 Main project page Previous entry Next entry
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Week 4 Day 4TSL - Agrobacterium-mediated transient expression in leaves of N. benthamiana PROTOCOL: 1. Single colony of each constructs 2,3,4,10,11 grown overnight at 28 °C in 5 ml media containing chloramphenicol. 2. Pellet by centrifugation at 4000 rpm for 15 min. 3. Resuspend in 10 mM MES, 10 mM MgCl2 and 150 μM Acetosyringone. 4. 1:100 dilution to give a final OD of 0.1-0.5 5. Incubate at RT for 4 hours. 6. Create a small puncture using a needle on the underside of N. benthamiana leaves of plants 6-8 weeks old. Using a blunt syringe infiltrate the agro constructs into the leaf puncture. Each construct is infiltrated into 4 sections on one leaf. 2 constructs per plant. 7. Store plants in controlled environment at 19-23 °C for 3 days.
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