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From 2012.igem.org

Results

• T7 Heme oxygenase (chloramphenicol): This biobrick assembly was successful. The plasmids will be sequenced and re-transformed into BL21 DE3 Rosetta & Star cells.
• Heme oxygenase (Kanamycin): Incubation is to be performed today and plasmid preparation using the Qiagen protocol will be performed. No colonies were present on the T7 minus heme oxygenase on ampicillin.
• Deinococcus: Plasmid sequencing will be performed and transformation was successful on kanamycin plates. The Deinococcus on the kanamycin was unsuccessful.
• T7 Agrobacterium: no colonies were present on the chloramphenicol plates.
• Agrobacterium: no colonies were present on either Kanamycin or Ampicillin plates.

	#1C	#2K	#2A	#3K	#3A	#4C	#5K	#5A
1. Addition of 20ng Gene Block Fragments in appropriate tube	2µl Hemo_T7A	2µl Hemo_A	2µl Hemo_A	2µl Deino_A	2µl Deino_A	2µl Agro_T7A	2µl Agro_A	2µl Agro_A
	2µl Hemo_B	2µl Hemo_B	2µl Hemo_B	2µl Deino_B	2µl Deino_B	2µl Agro_B	2µl Agro_B	2µl Agro_B
	nil	nil	nil	2µl Deino_C	2µl Deino_C	2µl Agro_C	2µl Agro_C	2µl Agro_C
	nil	nil	nil	2µl Deino_D	2µl Deino_D	2µl Agro_D	2µl Agro_D	2µl Agro_D
	nil	nil	nil	2µl Deino_E	2µl Deino_E	2µl Agro_E	2µl Agro_E	2µl Agro_E
2. Addition of 0.05 pmol of vector	2.7µl PSB-1C3	2.9µl PSB-1K3	2.8µl PSB-1A3	2.9 µl PSB-1K3	2.8 µl PSB-1A3	2.7µl PSB-1C3	2.9µl PSB-1K3	2.8µl PSB-1A3
3. Addition of Gibson Master Mix (µl)	10	10	10	12.9	12.8	12.7	12.9	12.8
4. Addition of deionised H2O	3.3µl	3.1µl	3.2µl	nil	nil	nil	nil	nil
Outcome