Team:Macquarie Australia/Protocols/GibsonAssembly

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Revision as of 10:16, 5 September 2012

Gibson Assembly Protocol

Before entering the lab, the assigned Gibson Assembly team for the day organised agar plate requirements, concentration calculations and equipment requirements.

Instruction	#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

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 {| border="2" cellpadding="0" cellspacing="0" align="center"
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-! scope="col"| Instruction
+! scope="col" colspan="1" | Instruction
 ! scope="col"| #1C
 ! scope="col"| #2K