Team:ULaVerne Collab/Notebook
Notebook
June
Week 2
(6/12) Restriction Digestion of PETase and K1357009 - Vector
(6/13) Restriction Digestion Old Vector K1357009 + pSB1C3 Ligation of PCR Purified Products Transformation of Ligation rxns
(6/14) Transformation Commercial Cells: 1. 1uL of Ligated Vector and Insert Non-Comm: 2. 1 uL Ligated Vector and Insert 3. 5 uL Ligated Vector and Insert 4. 1 uL Vector only 5. 5 uL Vector only
Week 3
(6/18) Overnight Colonies of Transformations Plan to pick 4+ colonies to screen ligations (more depending on how many colonies were on your control ligation plate.) Pick 2+ colonies when growing up bacteria for DNA stocks.
(6/19) Miniprep, Restriction Digest of Miniprep & GEL, Digestion of Promoter J23110
(6/21) Digestion Constitutive Promoter J23110, PCR Amplification of PETase
(6/22) GEL Purification, Restriction Digestion of PeTase and K1357009
Week 4
(6/25) Ligation, Transformation Results PeTase + promoter control 1 uL - 19 PeTase + promoter ligation 1uL - 30 PeTase + promoter control 5 uL - 40 PeTase + promoter ligation 5 uL - 54 Colonies PeTase +promoter 1 uL- 8 colonies PeTase + promoter 5 uL - 10 colonies
(6/26) Overnights Colonies of Transformation
(6/27) Miniprep
(6/29) Electroporation, Nanodrop
July
Week 1
(7/2) Gel Electrophoresis of Digested Vector w/ E + X, ElectroTransformation Data
Week 2
(7/12) Electrocompetent Cells 2, Digestion of PETase/ K1357009 from (5/29): Miniprep #2, #5, #6, NanoDrops of 5/29 Minipreps: PETase & K1357009, Gel Electrophoresis of PETase/ K1357009 from (5/29) and J23110 from Registry
Week 3
(7/16) Digestion of PETase/ K1357009 from (5/29): Miniprep #1 #4, #8
(7/17) Electro Competent Cells, Nanodrop of Gel Purification
(7/18) Ligation Calculation & Procedure, PCR Purification of Ligation Mixtures, ElectroTransformation, Mini Prep
(07/19) Chemical transformation of Ligations, Nanodrop PETase/ K1357009 *from (5/29)*: Miniprep #1 #4, Digestion (high DNA), Gel Electrophoresis, Gel Purification
(7/20) Ligation Reaction of J23110 Vector + PETase/K1357009
Week 4
(7/23) Interlab study, picking colonies from plates
(7/24) Miniprep, Nanodrops
(7/25) Miniprep, Nanodrop of Mini Prep PETase Circuit (Attempt 1), Digest of PETase Circuit (Attempt 1)
(7/26) Miniprep the Overnight, Nanodrop of MP, Digestions J23 and PET/K13
Week 5
(7/30) PCR Reaction, Primer Rehydration, Dilution of Promoter Stock, Promoter PCR, mHETase PCR, Modified Petase PCR for Cutinase test circuit, Modified Petase PCR for mHETase test circuit, Nanodrop
(7/31) Gibson Assembly Reaction 1 and PCR Reaction 2, Q5 2X PCR Reaction, Amil-CP Reaction (2), Curli Non Gibson PCR, OLD PETase PCR, Promoter Reactions
August
Week 1
(8/1) PCR products so far,
(8/2) PCR Purification of Curli Non-Gibson, Nanodrop Curli Non-Gibson, curli PCR for Gibson, Promoter PCR , Old PETase gel, PCR products
(8/3) Curli PCR Gibson redo sorry, Promoter Attempt 3,
Week 2
(8/7) Gibson Assembly, Cutinase Test circuit, PETase control circuit, mHETase test circuit, PCR Reaction, Cutinase for Old PETase,
(8/8) Gibson assembly of PETase mHet circuit,
(8/9) Mini Prep, Nanodrop of Gibson Mini Preps, Digestions Gibson Mini Preps (20 uL rxn)
(8/10) Mini Prep, Nanodrop oF Gibson Mini Preps, Digestions, Digestion #2
Week 3
(8/13) Digestion Gibson Mini Preps (20 uL rxn ), Gel, Pick colonies
(8/14) Nanodrop, miniprep of gibson parts, Digestions, Gel
(8/15) Nanodrop
Week 4
(8/20) PCR Tests, CUT mHet test CUT pcr, Old Pet CTRL PCR,
(8/21) Miniprep CUT test, nanodrop CUT test, PCR CUT Test CUT 1, CUT test CUT 2
September
Week 1
(9/4) Beginning of characterization
(9/7) Sodium Dodecyl Sulphate PolyAcrylamide Gel Electrophoresis
(09/08) Dilutions for Protein Gel
Week 2
(9/12) Sample Prep for Protein Gel
(9/15) International Coastal Cleanup
Week 3
(9/20) Sample Prep 2 for Protein Gel
(9/21) Staining and Destaining
Week 4
(9/25) Protein Extraction
(9/27) Math Modeling
October
Week 1
(10/1) Begin working on Wiki