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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== | ==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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Revision as of 06:48, 6 August 2014
 iGEM Project name 1
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6th August 2014
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
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