To scrutinize the results and properly understand the day to day progress of the Ovulaid team’s scientific work an exhaustive online notebook was kept, completely reflecting the team’s physical laboratory notebooks. The online notebook is structured by week and underneath you can see the experiments carried out that week. To see the details of each day and experiment, click on the week number.

Biosensor Notebook

Week 21 (20th-26th of May)
- CONSTRUCT AND PRIMER DESIGN
- DILUTION OF PRIMERS AND GENE FRAGMENTS
- NORMAL PCR AMPLIFICATION: X7-PFU POLYMERASE
- GEL ELECTROPHORESIS
- PURIFICATION OF PCR PRODUCT
Week 22 (27th of May-2nd of June)
- NANODROP
- ADAPTED USER REACTION (LIGATION)
- MAKING PLATES FOR E.COLI TO GROW
- E.COLI TRANSFORMATION
- COLONY PCR (E.COLI)
- GEL ELECTROPHORESIS
- TRANSFORMED E. COLI COLONIES INTO LIQUID CULTURE
- PLASMID DNA EXTRACTION
Week 23 (3rd-9th of June)
- SEQUENCING
- USER-LIGATION/E.COLI TRANSFORMATION PART 2: SAMPLE 4 & 3
- COLONY PCR
- GEL ELECTROPHORESIS
Week 24 (10th-16th of June)
- DNA EXTRACTION
- SEQUENCING OF SAMPLE S3C4 AND S3C5
- ADAPTED USER LIGATION SAMPLE 1, 2, 4, 6
Week 25 (17th-23rd of June)
- EXAM WEEK
Week 26 (24th-30th of June)
- COMPETENT CELLS
- NORMAL PCR AMPLIFICATION: X7-PFU POLYMERASE
- GEL ELECTROPHORESIS FOR SAMPLE 1, 11, 2 AND 12
- DNA PURIFICATION FOR SAMPLE 1, 2, 11 AND 12
- E. COLI TRANSFORMATION SAMPLES 1, 2, 4, 6
- MAKING OUR OWN USER VECTOR STOCK
- COLONY PCR FOR SAMPLES 1, 2, 4 AND 6
- COMPETENT CELL TEST
- GEL ELECTROPHORESIS FOR SAMPLES 1, 2, 4, AND 6
- PLASMID PURIFICATION
Week 27 (1st-7th of July)
- PCR AMPLIFICATION SAMPLE 1,2, 11 AND 12
- GEL ELECTROPHORESIS FOR SAMPLE 1, 2, 11 AND 12
- PLASMID DNA EXTRACTION
- E.COLI INCUBATION FOR VECTOR BACKBONE AMPLIFICATION
- PREPARING SAMPLES FOR SEQUENCING
- COMPETENT CELL TEST
- PLASMID PURIFICATION
- GEL ELECTROPHORESIS
- FIRST HUMAN LHCGR PCR AMPLIFICATION: X7-PFU POLYMERASE
- GEL ELECTROPHORESIS
Week 28 (8th-14th of July)
- 2nd PCR AMPLIFICATION: X7-PFU POLYMERASE AND GEL ELECTROPHORESIS
- SEQUENCING FOR GPER, GPER-sfGFP, XLHCGR and XLHCGR-sfGFP
- 3RD PCR AMPLIFICATION: X7-PFU POLYMERASE AND GEL ELECTROPHORESIS
- 4th PCR AMPLIFICATION: X7-PFU POLYMERASE AND GEL ELECTROPHORESIS
- YEAST TRANSFORMATION
- PCR AMPLIFICATION
Week 29 (15th-21st of July)
- COLONY PCR OF TRANSFORMED YEAST
- REDO OF YEAST COLONY PCR
- O/N CULTURES FOR GLYCEROL STOCKS
- GLYCEROL STOCKS
- PLASMID PURIFICATION
- PCR AMPLIFICATION
Week 30 (22nd-28th of July)
- YEAST TRANSFORMATION
- MEDIA AND REAGENT PREPARATION
- PRIMER DILUTION
- EZNA PLASMID PURIFICATION
- SAMPLE PREPARATION FOR SEQUENCING
- GLYCEROL STOCK
- EZNA PLASMID PURIFICATION
- GEL ELECTROPHORESIS OF OLD PCR PRODUCTS
- E. COLI TRANSFORMATION WITH Ass2A-pPGK1-GPA1-Gas
- COLONY PCR I OF TRANSFORMED YEAST FROM 22ND JULY 2019
Week 31 (29th of July-4th of August)
- GEL ELECTROPHORESIS OF E.COLI TRANSFORMATION CONSTRUCTS AND YEAST COLONY PCR SAMPLE
- COLONY PCR OF TRANSFORMED E. COLI
- COLONY PCR II OF TRANSFORMED YEAST FROM 22ND JULY 2019
- INOCULATION OF POSITIVE YEAST COLONIES OF CONSTRUCT X3A-pCCW12-GPER-sfGFP
- PLASMID PURIFICATION FOR Ass2A-pPGK1-GPA1-Gas EXPRESSING E. COLI
- FLUORESCENCE MICROSCOPE OF POSITIVE YEAST COLONIES
- PCR AMPLIFICATION of TPST1 & TPST2
- YEAST TRANSFORMATION
Week 32 (5th-11th of August)
- EZNA PLASMID PURIFICATION
- YEAST COLONY PCR
- COLONY PCR OF YEAST
- INOCULATION FROM PLATES
- USER LIGATION
- GEL ELECTROPHORESIS
- YEAST STREAKING
- GLYCEROL STOCKS
- PREPARATION OF OLD SAMPLES FOR SEQUENCING
Week 33 (12th-18th of August)
- USER LIGATION
- E. COLI TRANSFORMATION
- REPETITION OF USER LIGATION
- REPETITION OF E. COLI TRANSFORMATION
Week 34 (19th-25th of August)
- REPETITION OF COLONY PCR
- USER LIGATION
Week 35 (26th of August-1st of September)
- YEAST COLONY PCR
- GLU-U AGAR PREPARATION
- RUN OF GEL WITH COLONY PCR SAMPLE FROM 28/8
- PLASMID LIGATION
- INCUBATION OF YEAST CULTURES
- YEAST TRANSFORMATION
- GLYCEROL STOCKS
Week 36 (2nd-8th of September)
- YEAST COLONY PCR
- SETTING UP E.COLI CULTURES
- PROTEIN PLASMID PURIFICATION OF E.COLI CULTURES
- NANODROP
Week 37 (9th-15th of September)
- SETTING-UP CULTURES FOR BIOACTIVITY ASSAY
- FIRST ESTRADIOL BIOACTIVITY ASSAY
Week 38 (16th-22nd of September)
- SECOND ESTRADIOL BIOACTIVITY ASSAY
Week 39 (23rd-29th of September)
- Third Estradiol bioactivity assay
- Fluorescence Microscopy
Week 40 (30th of September-6th October)
- Fluorescence Confocal Microscopy
- Miniprep - Plasmid DNA purification
Week 41 (7th-13th October)
- Bioactivity assay
Week 42 (14th-20th October)
- 1st Western Blot with anti-GFP antibodies
- Confocal microscopy
- Repetition of the anti-sfGFP Western Blot

Kill Switch Notebook

Week 32 (5th-11th of August)
- PRIMERS AND G-BLOCKS DILUTION
- PCR AMPLIFICATION OF BAX AND BI-1
- GEL ELECTROPHORESIS
- PCR CLEAN UP
Week 33 (12th-18th of August)
- PCR AMPLIFICATION OF BI-1, GAL1, TEF1 AND ADH2
- GEL ELECTROPHORESIS OF PCR PRODUCTS
- PLASMID DIGESTION FOR USER LIGATION
- PCR CLEAN UP
- USER LIGATION
- E.COLI TRANSFORMATION
- COLONY PCR
- REPETITION OF COLONY PCR
Week 34 (19th-25th of August)
- GEL ELECTROPHORESIS OF COLONY PCR PRODUCTS
- PLASMID PURIFICATION
- PREPARATIONS FOR YEAST TRANSFORMATION
- PLASMID DIGESTION
- YEAST TRANSFORMATION
Week 35 (26th of August-1st of September)
- YEAST COLONY PCR
- GEL ELECTROPHORESIS FOR YEAST COLONY PCR
Week 36 (2nd-8th of September)
- SET UP A CULTURE OF THE X12C VECTOR
- GAL-INDUCTION ASSAY: MEDIA PREPARATION
- GAL-INDUCTION ASSAY: PREPARING PLATES
- GAL-INDUCTION ASSAY: PREPARE A CULTRE AND PLATE IT
- PURIFICATION OF X12C
Week 37 (9th-15th of September)
- SECOND GAL-INDUCTION ASSAY
- PLASMID DIGESTION
- SAMPLE DIGESTION FOR YEAST TRANSFORMATION
- SETTING UP YEAST FOR TRANSFORMATION
Week 38 (16th-22nd of September)
- YEAST COLONY PCR
- YEAST COLONY PCR
- STORING OF POSTIVE pGAL1-BAX CULTURE
Week 39 (23rd-29th of September)
- NO EXPERIMENTS
Week 40 (30th of September-6th of October)
- INOCULATION OF E.COLI CONTAINING pTEF-BI-1 & pGAL1-BI-1
Week 41 (7th-13th of October)
- Inoculation for gal-induction assay
- Sequencing
- Gal-induction assay (liquid) of pGAL1-BAX
- Preparing plates for gal-induction assay of the plasmid constructs
- Dual plasmid transformation
- Preparation of plates for the plasmid construct assays
- Miniprep
- Gal-induction assay of pGAL1-BAX
- Inoculation of yeast O/N cultures for WB
Week 42 (14th-20th of October)
- Restreak ADH2-BAX+GAL1-BI plasmid transformants
- Yeast transformation
- Restreaks from Monday were checked
- Gal-induction spot test of integrated and plasmid-expressed pGAL1-BAX
- Restreak of restreak was made