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

iGEM Project name 1

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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