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iGEM 2016 NUS SINGAPORE

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Wet Lab Notebook: RIOT RESPONDER

Jun Jul Aug Sep Oct

Jun

2016

Overall Strategies

1 June, 2016 Milestone 1 : Overall cloning design

12 June, 2016 Milestone 2: Construction of pConst-LuxR-HA

Construct 1:

Fig 1. A. LuxR gene amplified from iGEM distribution 2016 with designed primers GEM002 and GEM005. The expected band size of LuxR with overhang is 795 bp. B.LuxR stitiched with terminater (BBa_B0015) with overlapping PCR. C. Constructed full-length RBS-pConstitutive-LuxR-HA-Terminator with expected size of 1034 bp.

July

2016

Construct 2:

Fig 2. A. LuxI gene is cloned from iGEM2016 distribution kit with primers GEM008 and GEM011. The observed band size of LuxI with overhangs is close to the expected band size of 629 bp. B.Terminator BBa_B0015 is amplified using GEM009 and 012 and stitiched with LuxI. C. Promoter-RBS fragment is synthesized and stitiched to LuxI-terminator by PCR reaction. The observed band size for full LuxI construct is close to the expected band size of 936 bp.