Team:Exeter/Notebook
June
18-22/06/18: Pitched four project ideas, and Project Perchlorate was chosen.
Members: Team
26/06/18: Contacted LA Department of Water & Power, South West Water, SnapGene , Goonhilly Space Station, Airbus, and Eppendorf.
Members: Fiona
29/06/18: InterLab sign-up, project description & safety form submitted.
Members: Team
July
05/07/18: InterLab Calibrations 1, 2 and 3 .
Team: Amy, Emily, Ingebjørg, and Sophie
06/07/18: Interlab Cell Measurement Protocol attempt 1. Call with David Webster.
Team: Amy, Eleanor, Emily, Ingebjørg, Matt, and Sophie
10-11/07/18: Interlab Cell Measurement Protocol attempt 2.
Team: Amy and Sophie
12/07/18: University of Oxford UK iGEM meet-up attended. Water samples given to the University of Exeter.
Team: Amy, Ingebjørg, Matt, and Sophie
13/07/18: Contacted John Collins from synbiCITE.
Team: Matt
16/07/18: InterLab Cell Measurement Protocol attempt 3.
Team: Emily and Ingebjørg
17/07/18: Absorbance readings for InterLab study.
Team: Emily and Ingebjørg
19/07/18: Cell growth, sampling and assay technical report for InterLab. Flow cytometry. Contacted Nick Musgrove re: bioreactor. Received Illinois and Virginia water reports.
Team: Amy, Eleanor, Emily, Ingebjørg, Juliana, Matt, and Nahnsu
20/07/18: Colony Forming Units of E. coli for InterLab. Mark Strawbridge re: bioreactor.
Team: Amy, Matt, and Sophie
23/07/18: First competent cell preparation, one pot cloning with T7, sGFP constructs and B0015. Initial concept of bioreactor produced. Modelling work started.
Team: Eleanor, Fiona, Juliana, Nahnsu, and Ingebjørg
24/07/18: Preparation of competent cells. Transformation of competent cells with constructs from 24/07/18. Contacted Clive Butler.
Team: Eleanor and Matt
25/07/18: Competent cell preps. cld transformations started. Made up M9 media for cell growth testing.
Team: Eleanor, Matt, and Nahnsu
26/07/18: Further transformations of cld transformations. Copenhagen collaboration talk. Meeting with Professor Nicholas Harmer about enzyme modelling.
Team: Eleanor, Matt, and Ingebjørg
27/07/18: Contacted KCL and UCL for collaboration talks. Contacted Libby Jackson (UKSA). InterLab data submitted.
Team: Amy, Emily, Fiona, Ingebjørg, Matt, and Nahnsu
30/07/18: Cloning and transforming of the six superfolded sGFP constructs. Increasing rates theories and proof of concept. UCONN water report received.
Team: Eleanor, Emily, and Nahnsu
31/07/18: Cloning cld and making chloramphenicol agar plates. Preparation of overnight cultures using sGFP constructs from 30/7. Perchlorate solution curves.
Team: Eleanor, Emily, and Matt
August
01/08/18: sGFP DNA miniprep and Qubit assay.
Team: Eleanor
02/08/18: DNA prepared for sequencing submission. 24 hour OD curve of M9 solutions. Ben Reece water bioremediation call.
Team: Eleanor and Matt
03/08/18: cld miniprep. Bioreactor design sent to NASA (Melanie Pickett). Anaerobic incubation in 2% acetate solution.
Team: Eleanor, Sophie, and Matt
06/08/18: Overnights in 10% media solution. Ceri Lewis talk. Ben Reeve contacted again. More competent cells prepared.
Team: Matt and Eleanor.
07/08/18: Preparation of competent BL21(DE3) E. coli cells, inoculation of overnight cultures from 6/08, transformation of competent cells with cld and sGFP constructs. UCL collaboration talks continued. Alex Price contacted. Biodome placement investigated. Perchlorate assay produced.
Team: Eleanor, Matt, and Nahnsu
08/08/18: Enzyme digest of cld constructs. Copenhagen contacted. 24 hour OD curve of acetate solutions.
Team: Eleanor and Matt
09/08/18: Biosecurity researched. Plates and cld constructs transformed.
Team: Matt and Sophie
10/08/18: Collaboration with UC Davis. LB and acetate growth.
Team: Fiona and Matt
13/08/18: BL21 (DE3) E. coli transformations. Linear construct transformations. 24 OD curve of acetate and glucose.
Team: Eleanor, Fiona, and Matt
14/08/18: Overnight cultures prepared with Cld colonies.
Team: Eleanor
15/08/18: DTU contacted. PcrABCD digest, ligation, gel electrophoresis. Propagation and miniprep of cld/sGFP constructs in BL21 (DE3).
Team: Matt and Eleanor
16/08/18: cld and sGFP transformed into BL21 (DE3). Preparation of buffers for Fractionation of E. coli. Overnight cultures prepared for cld and sGFP constructs in BL21 (DE3) E. coli Melanie Pickett, Charles Cockrell contacted. Qubit assay. Transformation of pcrABCD complexes into DH5α E. coli.
Team: Eleanor, Emily, Matt, and Sophie
17/08/18: Propagation of cld and sGFP constructs into BL21 (DE3), fluorescence and absorbance of cultures measured using Tecan. Human practices refocused. Stanford-Brown talks.
Team: Eleanor, Fiona, Juliana, and Matt
20/08/18: BugBuster protein extraction reagent. Bolt bis-tris mini gel electrophoresis. Western blot. Contacted Professor Dussap.
Team: Eleanor and Matt
21/08/18: Visualisation of the Western Blot from 20/08. pcrABCD digest repeat, ligation and gel electrophoresis.
Team: Eleanor and Matt
22/08/18: Fractionation of sGFP constructs. iBind western system. sGFP periplasmic gel fraction. Alex Price contacted re: bioreactor.
Team: Eleanor and Matt
23/08/18: Visualisation of the Western Blot for the periplasmic fraction of the sGFP constructs from 22/08. Miniprep and qubit for cld/sGFP cds.
Team: Eleanor and Sophie
24/08/18: Protocol for making alginate beads determined. Track selections chosen and submitted.
Team: Matt and Team
26/08/18: CAD drawings of bioreactor.
Team: Matt
28/08/18: Andy Weir interview.
Team: Matt
29/08/18: Visited Bath to pick up bioreactor. Michael Curtis-Rowse call. Perchlorate assay finalised. Tecan perchlorate growth review.
Team: Amy, Fiona, Matt, Nahnsu, and Sophie
30/08/18: Cld assay solution, chlorite growth curve. Libby Jackson contacted again re: bioreactor.
Team: Juliana and Matt
31/08/18: Cld assay.
Team: Juliana
September
01/09/18: Bioreactor CAD drawings sent off.
Team: Matt
03/09/18: Met with Ewan Woodley. Cld activity in whole cell protocol.
Team: Fiona, Juliana, and Nahnsu
04-05/09/18: Overnights.
Team: Fiona, Juliana, and Sophie
07/09/18: Meeting with Paul Williams. Safety form submitted.
Team: Fiona, Nahnsu, and Sophie
10/09/18: Overnight cultures in preparation of oxygen testing protocol.
Team: Nahnsu and Sophie
11/09/18: Overnight cultures of wild type and pcrABCD construct prepared.
Team: Eleanor
12/09/18: Induction of overnight cultures from 11/09 with IPTG and growth. Optical density of oxygen testing protocol.
Team: Eleanor and Ingebjørg
13/09/18: Bolt gel electrophoresis and Western Blot of wild type and pcrABCD constructs from 12/09
Team: Eleanor
October
01/10/18: Western blots for PcrA, PcrC and PcrD performed. Double transformation pX1600 pcrABCD and cld into BL21(DE3) E. coli
Team: Eleanor, Dr. Mark Hewlett and Dr Paul James
03/10/18: Western blot for pcrABCD
Team: Dr. Mark Hewlett and Dr Chloe Singleton
04/10/18: pcrABCD successfully cloned into pSB1C3 plasmid
Team: Dr. Chloe Singleton
05/10/18: cld and pcrABCD successfully co-transformed into E.coli
Team: Dr. Paul James
12/10/18: Final Newcastle iGEM collaboration data received
Team: Newcastle iGEM team
14/10/18: Final Virginia iGEM collaboration data received
Team: Virgina iGEM team