IGEM:University of East Anglia (UEA), Norwich, UK/2009/Notebook/NRP-UEA-Norwich iGEM/2014/08/06: Difference between revisions

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(Autocreate 2014/08/06 Entry for IGEM:University_of_East_Anglia_(UEA),_Norwich,_UK/2009/Notebook/NRP-UEA-Norwich_iGEM)
 
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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"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
|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]]&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}}
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==Entry title==
==6th August 2014==
* Insert your content here.
* Set up level 2 dig-lig reactions (Vector, buffer, inserts, ligases)


Level 2 Constructs:


A= 35s_AvrBS3 + BS3_GFP


B= 35s_AvrBS3 + BS3_NbHb1
C= BS3_Bax + BS3_Bi1RNAi
D= 35s_AvrBS3 + BS3_Bax +BS3_BiRNAi
E= 35s_Bax1 + 35s_BiRNAi
F= PDF1.2_Bax1 + PDF1.2_BiRNAi
G=PR1_Bax + PR1_Bi1RNAi
* PCR of Level 2 Constructs (cycles: 3 cycles of
 10 minutes 37°C,
 10 minutes 16°C, 
followed by 
10 minutes 37°C, 
20 minutes 65°C
, 4°C forever)
*Made 1µl per ml Streptomycin plates. Spread with 100µl IPTG and 80µl of X-gal per plate
*Transformed 5µl of level 2 construct DNA from PCR into 50µl E.coli cells, electroporation of cells, spread (amount?) onto streptomycin plates, incubated overnight at 37oC
           
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Latest revision as of 00:09, 27 September 2017

iGEM Project name 1 Main project page
Previous entry      Next entry

6th August 2014

  • Set up level 2 dig-lig reactions (Vector, buffer, inserts, ligases)

Level 2 Constructs:

A= 35s_AvrBS3 + BS3_GFP

B= 35s_AvrBS3 + BS3_NbHb1

C= BS3_Bax + BS3_Bi1RNAi

D= 35s_AvrBS3 + BS3_Bax +BS3_BiRNAi

E= 35s_Bax1 + 35s_BiRNAi

F= PDF1.2_Bax1 + PDF1.2_BiRNAi

G=PR1_Bax + PR1_Bi1RNAi

  • PCR of Level 2 Constructs (cycles: 3 cycles of
 10 minutes 37°C,
 10 minutes 16°C, 
followed by 
10 minutes 37°C, 
20 minutes 65°C
, 4°C forever)
  • Made 1µl per ml Streptomycin plates. Spread with 100µl IPTG and 80µl of X-gal per plate
  • Transformed 5µl of level 2 construct DNA from PCR into 50µl E.coli cells, electroporation of cells, spread (amount?) onto streptomycin plates, incubated overnight at 37oC



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