Throughout the course of our project, we have continuously recorded our daily work, such as wetlab activities and
research findings. The notebook for Pneuumonsensor and Riboregulator are displayed separately.
Pneumosensor
June 2014
Week 1
June xx
Wet lab
·
·
·
·
June xx
Wet lab
·
·
·
·
June xx
Wet lab
·
·
·
·
June xx
Wet lab
·
·
·
·
Week 2
June xx
Wet lab
·
·
·
·
June xx
Wet lab
·
·
·
·
June xx
Wet lab
·
·
·
·
June xx
Wet lab
·
·
·
·
Week 3
June xx
Wet lab
·
·
·
·
June xx
Wet lab
·
·
·
·
June xx
Wet lab
·
·
·
·
June xx
Wet lab
·
·
·
·
Week 4
June 23
Wetlab
· Transformation of BBa_B0015 from iGEM Kit Plate 2011 and BBa_K880005
· Primer design for PCR of comX-myc and comW-FLAG from genomic DNA of S. Pneumoniae NCTC 7465 strain
·
·
June 24
Wetlab
· Inoculation of BBa_B0015
·
·
·
June 25
Wet lab
· Digestion of BBa_B0015 plasmid extraction product with XbaI and PstI, gel extraction and gel purification
· Inoculation of BBa_K880005
·
·
June 26
Wet lab
· Digestion of BBa_K880005 plasmid extraction product with SpeI and PstI, gel extraction and gel purification
·
·
·
June 27
Wet lab
· Ligation of BBa_B0015 insert cut with XbaI and PstI into pSB1C3-K880005 backbone cut with SpeI and PstI as negative control
·
·
·
Week 5
June 30
Wet lab
· Digestion of BBa_B0015 plasmid extraction product with XbaI and PstI, gel extraction and gel purification
· Gel check of purified BBa_B0015 insert
· Transformation of ligated pSB1C3-K880005.B0015
July 2014
Week 1
July 2
Wet lab
· Arrival of primers order for comX-myc and comW-FLAG
· Dilution of primers and design of experiment for both comX-myc and comW-FLAG
Inoculation of ligated pSB1C3-K880005.B0015
·
·
July 3
Wet lab
· PCR of comX-myc using Taq DNA Polymerase with ThermoPol Buffer to check whether primers are functional
· Gel check for comX-myc PCR product (PCR product present)
· PCR of comX-myc using Phusion High-Fidelity PCR Master Mix with HF Buffer
· Gel check for comX-myc PCR product (no PCR product present)
· Restriction digestion check on some candidate colonies of ligated pSB1C3-K880005.B0015 with XbaI and PstI
·
·
·
July 4
Wet lab
· PCR of comX-myc using Vent DNA Polymerase, PCR clean up
·
·
·
July 5
Wet lab
·
·
·
·
July 6
Wet lab
·
·
·
·
Week 2
July 7
Wet lab
· Digestion of comX-myc with XbaI and PstI, digestion clean up
· PCR of comW-FLAG using Vent DNA Polymerase, PCR clean up
· Dephosporylation of digested pSB1C3-K880005 backbone cut with SpeI and PstI
·
July 8
Wet lab
· Ligation of comX-myc cut with XbaI and PstI into pSB1C3-K880005 backbone cut with SpeI and PstI
· 2nd attempt of ligation of BBa_B0015 insert cut with XbaI and PstI into dephosporylated pSB1C3-K880005 backbone cut with SpeI and PstI as negative control
· Transformation of ligation products
·
·
·
July 9
Wet lab
· Inoculation of ligated pSB1C3-K880005.B0015 (2nd attempt) and ligated pSB1C3-K880005.comX-myc
·
·
·
July 10
Wet lab
· Another PCR of comW-FLAG using Vent DNA Polymerase, PCR clean up
· Restriction digestion check on some candidate colonies of ligated pSB1C3-K880005.B0015 (2nd attempt) and ligated pSB1C3-K880005.comX-myc, no insert wanted present
·
·
·
July 11
Wet lab
· Digestion of comW-FLAG with XbaI and PstI, digestion clean up
·
·
·
Week 3
July 14
Wet lab
· Ligation of comW-FLAG cut on XbaI and PstI into dephosporylated pSB1C3-K880005 backbone cut on SpeI and PstI
· Transformation of ligation products
· Inoculation of ligated dephosporylated pSB1C3-K880005.comX-myc
·
·
·
July 15
Wet lab
· Digestion check of ligated pSB1C3-K880005.comX-myc
· Digestion of pSB1C3-K880005.comX-myc on SpeI and PstI, but no insert wanted present
· PCR of comX using Vent DNA Polymerase, PCR clean-up
· Colony PCR of pSB1C3-K880005.comX-myc using Taq DNA Polymerase
· Gel check of pSB1C3-K880005.comX-myc PCR product (no PCR product present)
· Inoculation of dephosporylated pSB1C3-K880005.comX-myc, dephosporylated pSB1C3-K880005.comW-FLAG
·
·
·
·
July 16
Wet lab
· Digestion check of ligated pSB1C3-K880005.comW-FLAG
· Colony PCR of pSB1C3-K880005.comW-FLAG
· Gel check of pSB1C3-K880005.comW-FLAG PCR product (no PCR product present)
· PCR of comW using Vent DNA Polymerase (no PCR product present)
· Digestion of comX on XbaI and PstI
·
·
·
·
·
July 17
Wet lab
· PCR of comX-myc and comX using Vent DNA Polymerase, PCR clean-up
· PCR of comW using Vent DNA Polymerase
· Gel extraction and gel purification of comW PCR product
· Dephosporylation of BBa_K880005
·
·
·
·
July 18
Wet lab
· PCR of comX using Vent DNA Polymerase, PCR clean-up
· Digestion of comX PCR product on XbaI and PstI, digestion clean-up
· Digestion of comX-myc and comW PCR products, digestion clean-up
· Gel extraction and gel purification of digested comX and comX-myc
·
·
·
Week 4
July 21
Wet lab
· Gel purification of comX and comX-myc
· Ligation of comX-myc into dephosporylated pSB1C3-K880005 backbone with 1:4, 1:7, 1:10 ratio
· Ligation of comW-FLAG into dephosporylated pSB1C3-K880005 backbone with 1:4, 1:7, 1:10 ratio
· Ligation of comX into dephosporylated pSB1C3-K880005 backbone with 1:3 and 1:5 ratio
· Ligation of comW into dephosporylated pSB1C3-K880005 backbone with 1:4, 1:7, 1:10 ratio
· Colony PCR of pSB1C3-K880005.comW-FLAG using Taq DNA Polymerase
·
·
·
·
July 22
Wet lab
· Colony PCR of comX, comW, comX-myc, comW-FLAG and use BBa_J04450 as the positive control using Taq DNA Polymerase (no PCR product present)
· Inoculation of comX, comW, comX-myc, comW-FLAG
·
·
·
·
·
July 23
Wet lab
· Digestion check of comX, comW, comX-myc, comW-FLAG colony PCR products on XbaI and SpeI
· Inoculation of comW-FLAG
·
·
·
·
·
·
July 24
Wet lab
· Digestion check of pSB1C3-K880005.comW-FLAG on XbaI and SpeI-HF
· Ligation of comW-FLAG into dephosporylated pSB1C3-K880005 backbone
· Ligation of comX-myc into dephosporylated pSB1C3-K880005 backbone
· PCR of comX-myc and comX using Vent DNA Polymerase, gel extraction and gel purification
· Digestion of comX-myc and comX on XbaI and PstI-HF
·
·
·
·
July 25
Wet lab
· Colony PCR of comX-myc using Taq DNA Polymerase
· Gel check of comX-myc PCR product (no PCR product present)
·
·
·
·
·
Week 5
July 28
Wet lab
· Colony PCR of comX-myc using Taq DNA Polymerase
· Colony PCR of comw-FLAG using Taq DNA Polymerase
· Inoculation of comX-myc and comW-FLAG
·
·
·
·
July 29
Wet lab
· Digestion check of comX-myc and comW-FLAG on XbaI and PstI-HF
· Transformation of comX-myc and comW-FLAG
·
·
·
·
·
July 30
Wet lab
· Digestion check of comX-myc and comW-FLAG
· Inoculation of comX-myc and comW-FLAG
· Streaking plates of comX-myc and comW-FLAG
·
·
·
·
·
July 31
Wet lab
· Digestion check of comX-myc and comW-FLAG on XbaI and PstI-HF
· Redigest comW-FLAG on XmnI and NcoI
· PCR sequencing for comW and comX
· Ethanol precipitation for comW and comX
·
·
·
·
August 2013
Week 1
Aug xx
Wet lab
·
·
·
·
Aug xx
Wet lab
·
·
·
·
Aug xx
Wet lab
·
·
·
·
Aug xx
Wet lab
·
·
·
·
Week 2
Aug xx
Wet lab
·
·
·
·
Aug xx
Wet lab
·
·
·
·
Aug xx
Wet lab
·
·
·
·
Aug xx
Wet lab
·
·
·
·
Week 3
Aug xx
Wet lab
·
·
·
·
Aug xx
Wet lab
·
·
·
·
Aug xx
Wet lab
·
·
·
·
Aug xx
Wet lab
·
·
·
·
Week 4
Aug xx
Wet lab
·
·
·
·
Aug xx
Wet lab
·
·
·
·
Aug xx
Wet lab
·
·
·
·
Aug xx
Wet lab
·
·
·
·
Week 5
Aug xx
Wet lab
·
·
·
·
Aug xx
Wet lab
·
·
·
·
Aug xx
Wet lab
·
·
·
·
Aug xx
Wet lab
·
·
·
·
September 2013
Week 1
Sept xx
Wet lab
·
·
·
·
Sept xx
Wet lab
·
·
·
·
Sept xx
Wet lab
·
·
·
·
Sept xx
Wet lab
·
·
·
·
Sept xx
Wet lab
·
·
·
·
Week 2
Sept xx
Wet lab
·
·
·
·
Sept xx
Wet lab
·
·
·
·
Sept xx
Wet lab
·
·
·
·
Sept xx
Wet lab
·
·
·
·
Sept xx
Wet lab
·
·
·
·
Week 3
Sept xx
Wet lab
·
·
·
·
Sept xx
Wet lab
·
·
·
·
Sept xx
Wet lab
·
·
·
·
Sept xx
Wet lab
·
·
·
·
Sept xx
Wet lab
·
·
·
·
Week 4
Sept xx
Wet lab
·
·
·
·
Sept xx
Wet lab
·
·
·
·
Sept xx
Wet lab
·
·
·
·
Sept xx
Wet lab
·
·
·
·
Sept xx
Wet lab
·
·
·
·
Week 5
Sept xx
Wet lab
·
·
·
·
Sept xx
Wet lab
·
·
·
·
Sept xx
Wet lab
·
·
·
·
Sept xx
Wet lab
·
·
·
·
Sept xx
Wet lab
·
·
·
·
Riboregulator
June 2013
Week 1
June xx
Wet lab
·
·
·
·
June xx
Wet lab
·
·
·
·
June xx
Wet lab
·
·
·
·
June xx
Wet lab
·
·
·
·
Week 2
June xx
Wet lab
·
·
·
·
June xx
Wet lab
·
·
·
·
June xx
Wet lab
·
·
·
·
June xx
Wet lab
·
·
·
·
Week 3
June xx
Wet lab
·
·
·
·
June xx
Wet lab
·
·
·
·
June xx
Wet lab
·
·
·
·
June xx
Wet lab
·
·
·
·
Week 4
June xx
Wet lab
·
·
·
·
June xx
Wet lab
·
·
·
·
June xx
Wet lab
·
·
·
·
June xx
Wet lab
·
·
·
·
Week 5
June xx
Wet lab
·
·
·
·
June xx
Wet lab
·
·
·
·
June xx
Wet lab
·
·
·
·
June xx
Wet lab
·
·
·
·
July 2013
Week 1
July xx
Wet lab
·
·
·
·
July xx
Wet lab
·
·
·
·
July xx
Wet lab
·
·
·
·
July xx
Wet lab
·
·
·
·
Week 2
July xx
Wet lab
·
·
·
·
July xx
Wet lab
·
·
·
·
July xx
Wet lab
·
·
·
·
July xx
Wet lab
·
·
·
·
Week 3
July xx
Wet lab
·
·
·
·
July xx
Wet lab
·
·
·
·
July xx
Wet lab
·
·
·
·
July xx
Wet lab
·
·
·
·
Week 4
July xx
Wet lab
·
·
·
·
July xx
Wet lab
·
·
·
·
July xx
Wet lab
·
·
·
·
July xx
Wet lab
·
·
·
·
Week 5
July xx
Wet lab
·
·
·
·
July xx
Wet lab
·
·
·
·
July xx
Wet lab
·
·
·
·
July xx
Wet lab
·
·
·
·
August 2013
Week 1
Aug xx
Wet lab
·
·
·
·
Aug xx
Wet lab
·
·
·
·
Aug xx
Wet lab
·
·
·
·
Aug xx
Wet lab
·
·
·
·
Week 2
Aug xx
Wet lab
·
·
·
·
Aug xx
Wet lab
·
·
·
·
Aug xx
Wet lab
·
·
·
·
Aug xx
Wet lab
·
·
·
·
Week 3
Aug xx
Wet lab
·
·
·
·
Aug xx
Wet lab
·
·
·
·
Aug xx
Wet lab
·
·
·
·
Aug xx
Wet lab
·
·
·
·
Week 4
Aug xx
Wet lab
·
·
·
·
Aug xx
Wet lab
·
·
·
·
Aug xx
Wet lab
·
·
·
·
Aug xx
Wet lab
·
·
·
·
Week 5
Aug xx
Wet lab
·
·
·
·
Aug xx
Wet lab
·
·
·
·
Aug xx
Wet lab
·
·
·
·
Aug xx
Wet lab
·
·
·
·
September 2013
Week 1
Sept xx
Wet lab
·
·
·
·
Sept xx
Wet lab
·
·
·
·
Sept xx
Wet lab
·
·
·
·
Sept xx
Wet lab
·
·
·
·
Sept xx
Wet lab
·
·
·
·
Week 2
Sept xx
Wet lab
·
·
·
·
Sept xx
Wet lab
·
·
·
·
Sept xx
Wet lab
·
·
·
·
Sept xx
Wet lab
·
·
·
·
Sept xx
Wet lab
·
·
·
·
Week 3
Sept xx
Wet lab
·
·
·
·
Sept xx
Wet lab
·
·
·
·
Sept xx
Wet lab
·
·
·
·
Sept xx
Wet lab
·
·
·
·
Sept xx
Wet lab
·
·
·
·
Week 4
Sept xx
Wet lab
·
·
·
·
Sept xx
Wet lab
·
·
·
·
Sept xx
Wet lab
·
·
·
·
Sept xx
Wet lab
·
·
·
·
Sept xx
Wet lab
·
·
·
·
Week 5
Sept xx
Wet lab
·
·
·
·
Sept xx
Wet lab
·
·
·
·
Sept xx
Wet lab
·
·
·
·
Sept xx
Wet lab
·
·
·
·
Sept xx
Wet lab
·
·
·
·
|
"
