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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 (11/09/2012)	4 plasmids to sequence			2 plasmids to sequence		No colonies