January-May

January

• A box of pipette tips could ruin your life.
• Applications for new iGEM team members are finalized and approved.
• The new team meets for the first time!

Febuary

• My family is really big on suing
• Learning about circuit design
• Attending University lectures on synthetic biology
• Brainstorming topics for the project

March

• I’m dropping you off on the side of the road on the way home.
• Planning fundraising events
• Applying for University sponsored research grants
• Lab Training
• Learning about primer design

April

• We’re not poor, we’re environmentally conscious
• Project Proposal Presentations
• Choose final project topic
• Researching Sensing systems in E. coli
• Researching laboratory protocols
• Writing to companies to acquire sponsorships

May

• Oh dear, we made it mad
• Meet with Dr. Balaszi and Dr. Gergen to finalize project topic
• Making lists of supplies to purchase
• Literature research
• May 8th: That’s my life goal...to be an old person.
  • Meet with Dr. Balaszi for help with math modeling and tripeptide expression
  • Sequence tripeptides in a string instead of one at a time
  • Look for a protease to cut tripeptide repeats
• May 25th: If I could pull a rabbit out of a graduated cylinder...
  • Apply to volunteer and teach synthetic biology at the Long Island Maker Festival
• May 26th: Avoid breathing in smelly things.
  • First Day in the Laboratory!
  • Setting up the lab equipment
  • Researching a new protease
  • Finding a plasmid backbone
  • Literature research to modify glucose sensing system in E. coli with ompR mutants
  • Setting up controls to test each step of the procedure
  • Literature research to find protein export system
• May 27th: It’s like a pleasant cup of hot chocolate.
  • Finalizing Circuit constructs
  • Literature research to optimize sensor
  • Use of his tag to track sequences
  • Finding existing BioBricks to use in circuit
  • Working on presentation for Maker festival
• May 28th: The worst things that humanity has invented are genocide, the atomic bomb and black licorice.
  • Refining shopping list
  • Finalizing circuit
  • Working on presentation for Maker Festival
• May 29th: It’s kind of redundant having both of you on the team.
  • Learning how to BioBrick Melittin via Add-on-PCR
  • Literature research on rare codons
  • Meeting with Dr. Gergen to talk about how to detect tripeptides
  • Literature research on new protein export system

June

• June 1st: I need an adult
  • Literature research on another new protease
  • Find new plasmid for constructs
  • Literature research on bacteria recombination effects on constructs
  • Email Stony Brook professors for help detecting tripeptides
• June 2nd: It’s not going to be hard, it’s going to be very difficult
  • Literature research on HlyA secretion tag
  • Literature research on E. coli strains with T7 Polymerase integrated into genome
  • Meeting with Dr. Thanassi to learn about HlyA tag system
• June 3rd:
  • Decide on plasmid to use from iGEM kit
  • Order E.coli from Yale CGSC
  • Team presented project design at Wantagh High School
  • Video call with Synbiota for sponsorship
  • Emailed director of Humanology project for potential partnership
• June 4th: Lime green is not for science.
  • Meeting with Dr. Thanassi to discuss secretion system
  • Finalize two plasmid system with secretion system and tripeptides/protease
• June 5th: duh, diabetes
  • Literature research on linker to get secreted protease to outer membrane
• June 8th: I don’t trust anything anymore
  • Find new transporter for tripeptides
  • Looking for compatible plasmids in the two plasmid model
• June 9th: Everyone… drink the peptide (we didn't drink any peptides, safety first!)
  • Research outer membrane fractionation
  • Literature research on plasmid compatibility
• June 10th: I like dark blue not an azure
  • Find sequence for new protease
  • Found compatible plasmid
  • Ordered E. coli strains and plasmids
• June 11th: They’ll probably just drop an elephant on it.
  • Meeting with Dr. Gergen to finalize circuit design
  • Meeting with Dr. Karzai to finalize circuit design
  • Working on poster for Maker Festival
• June 12th: Now address me as the Buddha. Thank you.
  • Ordered IDT Constructs
  • Meet with Dr. Carlson, head of endocrinology at Stony Brook University Hospital, to talk about the medical ethics and applications of our project
• June 14th: That’s not how cancer works.
  • iGEM presents at the Long Island Maker Festival!
• June 17th: I’m not stealing his brain, it’s technically mine, he just didn’t die…
  • Making media
• June 18th: I can fall asleep doing anything, any time…
  • iGEM bake sale
  • E. coli cells arrive!
  • Plated cells to grow
• June 19th: It’s not like they are going to throw us in science jail…
  • Find Competent Cell Protocol
• June 22nd: Imagine a doctor giving you a drink that tastes good, but you know it’s actually poop.
  • Making competent cells
  • Literature research: Protease ATP requirements
• June 23rd: For future reference, I like blue gloves because they are sexier than purple ones.
  • Competent cell test
  • Learn to Measure OD of Liquid Culture
  • Literary Research of Secretion System
• June 24th: If I die in this lab, I expect one of you to reanimate me.
  • Resuspending IDT constructs
  • Testing Competent cells
  • Literary Research of Secretion System
• June 25th: You’d probably just vomit everywhere.
  • Digesting constructs
  • Testing Competent cells
• June 26th: No! Get some oil and light it on fire!
  • Tested Competent cells, learned not enough chloramp was used
• June 29th: Imagine Greg and I sober…
  • Making Competent cells
  • Testing Competent Cells (chloramp good, experimental not)
  • Learned to Run a Gel
• June 30th: Smile with your mistake!
  • Digesting constructs with new protocol
  • Competent cells test
  • Meeting with Dr. Czaplinski to troubleshoot competent cell problems
  • Went to molecular cloning facility to order primers

July

• July 1st: So lunch, then cry.
  • Presenting to Green Genes camp at Cold Spring Harbor Laboratory
  • Competent cells test using controls from Dr. Czaplinski
  • Troubleshooting transformation protocol
  • Meeting with Dr. Seeliger to discuss E.coli membrane compositions and sensing ability
• July 2nd: Oh shoot, he wants to be an oncologist! I’m going to be a fire fighter!
  • Making competent cells again, new protocol
  • Vector digest
  • Competent cells test
• July 3rd: I’m going to name my kid Kevin… especially if it’s a girl.
  • Competent Cell test success!
• July 6th: We’re a pretty fertilized species.
  • Start minipreps to extract transformed iGEM vector to use in digestion
  • Vector digest
  • Ligation (test 2 hours and overnight)
  • Gel extraction of Digested Vector
  • Literary Research into Origin Compatibility
• July 7th: Wow! It wasn’t the right math but it was on forever!
  • Vector Ligation, new protocol
  • Test Rubidium Competent Cells
• July 8th: They’re gonna die. Don’t get attached.
  • Presenting to Green Genes and DNA Science camp at Cold Spring Harbor Laboratory
  • PCR of Construct 1 (12 cycles- incorrect)
  • Transformations to test Rb competency, plate 4, and ligation product
• July 9th: You know when you see something hot and you can see the wiggles?
  • New Digestion protocol
  • More vector and construct digestions
  • Meeting with Dr. Gergen to discuss protocols
  • Meeting with Dr. Balaszi to discuss E. coli
• July 10th:
  • Digest without BSA
• July 13th: I identify most as a spatula.
  • Testing restriction enzymes
  • Obtaining new Xbal
  • Ordered more IDT constructs
  • More digestions
• July 14th: I’m no better than a monkey in a lab coat.
  • More digestions of construct
  • Ligation of Constructs
  • Finding a new protease
  • Literature research: diseases that can be treated with tripeptides
• July 15th:
  • More digestions of construct
  • Successful PCR of constructs
  • Teaching at Cold Spring Harbor
  • Literary research on Diabetes Insipidous
• July 16th: If Greg doesn’t eat before 1 he must feast on the blood of humans.
  • More construct digestions
  • New ligation protocol
  • Troubleshooting vector digestion
• July 17th: Manslaughter? Manslaughter.
  • More digestions and ligations
• July 20th: The shapes add a little bit of fun to it.
  • More digestion and ligation
  • Exploring ways to make gels of a higher resolution
• July 21st: Can we practice flicking?
  • More digestions and ligations
• July 22nd: Cells are so stupid.
  • More digestions and ligations
  • Meeting with Dr. Czaplinski to troubleshoot ligation protocol
• July 24th: He needs to know how heavily he’s vomiting.
  • Finding concentration of DNA using a gel
  • More digestion and ligations
  • Making Competent cells
• July 27th: He’s bald
  • More digestions and ligations
  • Troubleshooting gel extraction protocol
  • Successful ligation and transformation of construct 3!
• July 28th: Motherhood made me soft.
  • More digestions and ligations
  • Working on URECA poster
  • Modifying Melittin BioBrick sequence
  • Wiki content writing
• July 29th: One thing goes wrong and they’re my children, not our children
  • More PCR
  • More digestions and ligations
  • Register for URECA summer research symposium
  • Working on URECA poster
• July 30th: Either you succeed in life or you’re worthless to me.
  • Successful transformation of construct 3
  • Sequence construct 3
  • Archaea collaboration with UGA
  • Refining project application
  • Working on project abstract
  • Looking at alternative protesases
• July 31th: If I make a hole big enough for bread to go through, that doesn’t mean that people can.
  • Successful PCR

August-September

• August 3rd: I could never wear pants in lab
  • More digestions and ligations
  • Research potential applications of project in microflora and microcapsules
• August 4th: Why is this our problem? -Because we have so many, we like to focus on the little ones
  • More digestions and ligations
• August 5th: It’s like giving birth to words.
  • More digestions and ligations
  • Testing tripeptide production in construct 3 via SDS page gel
  • Testing inducibility of tripeptide production
• August 7th: I’m the least arrogant at 11:50pm because I’ve used it all up
  • More digestions and ligations
  • Presenting at URECA summer research symposium
  • Finalize project title and submit abstract
• August 10th: Or, depending on your view of life, best case scenario: we all die
  • Troubleshoot oligo resuspension
  • Resuspend oligos
  • More digestions and ligations
• August 11th: I like it when it’s wet outside, it’s like an extra shower
  • Troubleshooting ligation protocol
  • More digestions and ligations
  • SDS page gel to test tripeptide production
• August 12th: My mood gets so good when I look at myself
  • Presenting to DNA Science at Cold Spring Harbor Laboratory
  • Troubleshooting transformation
  • Successful PCR of constructs
  • More digestions and ligations
• August 13th: You have to pay extra for the bathroom
  • Successful transformation of melittin mCherry optimus
  • More digestions and ligations
• August 14th: I’m like that transformed E. coli cell that actually transformed.
  • More digestions and ligations
  • Successful transformation of Construct 5 and melittin, melittin optimus and melittin mCherry
  • Meeting with the Dean of the College of Arts and Sciences
• August 15th: Hold your pants
  • More digestions and ligations
• August 17th: Is it sketchy? Cuz’ I’ll get mugged or something
  • Sequencing constructs
• August 19th: It’s animated -Yes, because the real dinosaurs are hard to book
  • Presenting to Green Genes camp at Cold Spring Harbor Laboratory
  • Attending training session at New York Hall of Science
• August 23th: We can put diabetes medicine in your eye
  • Presenting at the New York Hall of Sciences synthetic biology event

September

What if we get arrested in Boston?
• Back to school
• More digestions and ligations
• BioBricking
• Testing constructs
• Working on team poster and presentation
• Writing wiki content