Welcome to our notebook, the log of all our hard work this summer! Below are the summaries of what occured each week; if you would like to see more detail, click on the weeks to expand it.
May 30-June 2
‣ Met with our advisors and worked on construct design for this week
‣ Set up the lab for start of wet lab work
June 3-9
‣ Received our cyanobacteria and started culturing them
‣ Started a growth experiment to determine ideal growing conditions in our incubators (1 paper vs 3 sheets of paper)
June 10-16
‣ Continuation of growth experiments for cyanobacteria
‣ Spectrophotometry measurements for growth experiment
‣ Primers design for PCR and PCR information
‣ Spectrophotometric measurements to determine growth curve of cyanobacteria
June 17-23
‣ Continuation of spectrophotometry measurements for growth curve
‣ Started work for E. coli plasmid cultures by making agar plates
‣ Tested antibiotic solutions (streptomycin and spectinomycin) on plasmids
‣ Miniprep of 1579 DNA
June 24-30
‣ Continuation of spectrophotometry measurements
‣ Splitting/growing and culture of cyanobacteria
‣ Growing up plasmids received from Addgene
‣ Constructs from IDT arrived and primers for PCR arrived
‣ Both constructs and primers were resuspended
‣ PCR and PCR purification of all constructs that arrived
‣ Ran multiple gels on plasmid cultures and tested restriction enzymes in digest
‣ Start of Interlab Calibration 1/Calibration 2
July 1-7
‣ Continuation of spectrophotometry measurements
‣ Splitting/growing and culture of cyanobacteria
‣ Gel extraction and gel purification for plasmids
‣ Interlab - Finished Calibration 1/2/3
‣ Made chloramphenicol stock
‣ PCR and gel purification of constructs
July 8-14
‣ PCR, gel extraction and gel purification of constructs
‣ Gel purification of plasmids
‣ Made glycerol stock of plasmids and frozen stocks of cyanobacteria
‣ Splitting/growing and culture of cyanobacteria
‣ Started Interlab transformation and inoculation of E. coli (Day 2)
July 15-21
‣ Splitting/growing and culture of cyanobacteria
‣ Finished interlab inoculation of E. coli colonies (Day 2)
‣ Gel extraction and purification of constructs (troubleshooting)
‣ PCR adjustments and trouble shooting for constructs
‣ Miniprep, RE digest and DNA purification of plasmids
‣ Starting work with CO2 incubator for growth of cyanobacteria
‣ Obtained light data on CO2 for the incubator
July 22-28
‣ Splitting/growing and culture of cyanobacteria
‣ Interlab Day 3 - colony forming units protocol
‣ Contamination experiments for cyanobacteria
‣ PCR of constructs and ran products on gel
‣ First HiFi transformation of PCR product for cpc (with positive and negative controls)
July 29-August 4
‣ Splitting/growing and culture of cyanobacteria
‣ PCR of constructs and running them on a gel
‣ PCR purification of products
‣ HiFi of cpc, cpc 560, lone cscB, idiA, optimized EYFP and original EYFP
‣ Inoculated colonies with HiFi
‣ Made BG-11 plates for culture of transformed cyanobacteria
‣ Transformation of cyanobacteria with lone cscB, cpc, cpc 560
‣ PCR troubleshooting
‣ Gel purification troubleshooting
‣ Sodium bicarbonate growth experiments
August 5-11
‣ Sodium bicarbonate growth experiments
‣ Gel purification troubleshooting
‣ Splitting/growing and culture of cyanobacteria
‣ Cyanobacteria transformation of rbc
‣ HiFi for idiA, EYFP and positive controls
‣ PCR troubleshooting
August 12-18
‣ Sodium bicarbonate growth experiments
‣ Splitting/growing and culture of cyanobacteria
‣ Transformed and plated HiFi for Opto EYFP and orig. EYFP
‣ Restreaked cyanobacteria plates
‣ Transformation of idiA, psba2, rbc
‣ Miniprep and inoculation for biobricks
‣ Start of sucrose assay experiments
August 19-25
‣ Sodium bicarbonate growth experiments
‣ Splitting/growing and culture of cyanobacteria
‣ Sucrose assay experiments
‣ Biobrick miniprep
‣ Colony PCR
‣ Transformation of cyanobacteria
August 26-September 1
‣ Cyanobacteria sucrose assays with cscB
‣ PCR and PCR purification for biobrick group
‣ Transformed cyanobacteria with psbA2 and plated transformed bacteria
September 2-8
‣ Digestion of pSB1C3 and ligation with constructs
‣ Transformation of ligated cpc560
‣ Inoculation of cpc560, idiA
‣ Made BG-11 media for inoculations
September 9-15
‣ Miniprep of transformed cpc560
‣ Inoculation of psbA2 colonies for cyanobacteria
‣ Ligation and Transformation of cpc for biobrick
‣ Luciferase assays for cpc
‣ Making LB and CAM plates for biobrick
‣ Inoculation of cpc
‣ Sucrose assay for cscB
September 16-22
‣ Miniprep of idiA, cpc
‣ PCR of cscB, psbA2 and PCR purification
‣ Ligation and transformation of psbA2
‣ Another luciferase experiment
September 23-29
‣ Sequencing for biobrick DNA for cpc560, orig EYFP, opto EYFP, cpc, idiA
‣ Sucrose experiment for cscB
September 30-October 6
‣ RE digest of backbone, psbA2, cpc560, idiA, cscB
‣ Transformation of psbA2, cpc560, idiA, cscB
‣ Inoculation of transformed colonies
‣ Miniprep of inoculation and sequencing
‣ Sucrose assay with cscB
‣ Luciferase assay for cpc, cpc560, psbA2
October 7-13
‣ Miniprep of transformed biobrick colonies for opto EYFP, cscB, psbA2, idiA, cpc560
‣ Dried DNA and submitted to iGEM HQ
‣ Sucrose assay with cscB and increased amount of IPTG
‣ Luciferase experiment with psbA2
October 14-17
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The Stony Brook iGEM Team is proud to present to you their sweet and energy filled project! Made with love <3
Email: igem.sbu@gmail.com
