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Experiment Timeline


18th June

  • Preparation of LB medium
  • Amplification of parts from registry (pTet, TetR, HO1, Low copy plasmid)

19th June

  • Antibiotic stock preparation
  • Inoculation

20th June

  • Our first Plasmid Extraction (PE)!

22nd June

  • First Double Digestion (DD) and Gel Check!

23rd June

  • PE & DD of the registry constructs….

24th June

  • Our direct synthesized parts finally arrived. We now have CHE (cholesterol esterase), IFP(Infrared protein 1.4) and pNO (NO sensing promoter).

26th June

  • PE , DD and Gel check for the direct synthesized gene sequence.

27th June

  • Gel Check for CHE, IFP and pNO


1st July

  • PE for plaqIQ, J23101, J23119, B0034(rbs), B0015(Term-T for short), B0034-GFP-B0015

2nd July

  • PE for pNO, IFP, CHE
  • DD of pNO, J23119, J23101, rbs-GFP-T
  • Our first ligation!

3rd July

  • DD of CHE, IFP and B0015, plaqIQ and plux.
  • Ligation.

4th July

  • PE: pNO-GFP; J23101-GFP; J23119-GFP; HO1
  • DD: HO1

7th July

  • PE: CHE, IFP, placIQ
  • DD:CHE-T, IFP-T. HO1, Term
  • Ligation

8th July

  • DD: pNO-GFP; J23101-GFP; J23119-GFP

9th July

  • PE
  • DD: ptet. r-CHE-T, r-HO1, r-IFP-T
  • Ligation: r-HO1 +r-IFP-T

10th July

  • PE
  • DD: ptet; r-CHE-T, Term, tetR

11th July

  • PE
  • DD: r-CHE-T; r-HO1-r-IFP-T;CHE-T; r-CHE-T

13th July

  • DD: ptet; r-CHE-T; r-HO1-r-IFP-T; TetR; Term
  • Ligation: ptet + r-CHE-T; ptet + r-HO1-r-IFP-T; TetR + Term

15th July

  • PE
  • DD: pTet; rbs; IFP; CHE; TetR-T

16th July

  • DD: pTet; r-CHE-T; Inv; r-HO1-r-IFP-T; Term; TetR
  • Ligation

18th July

  • PE
  • DD and Single Digestion (SD) :r-HO1-r-IFP-T (because they have the same size)

20th July

  • DD:pTet; J23119; r-HO1-r-IFP-T; TetR; Term; placIQ; inv
  • Ligation: pTet +r-HO1-r-IFP-T; J23119 + r-HO1-r-IFP-T; placIQ +Inv; TetR +Term
  • PE

23rd July

  • PE
  • DD:J23119; Inv; r-HO1; r-GFP-T; pTet; placIQ-Inv; TetR-T

24th July

  • PE
  • DD: J23119; r-CHE-T; pTet; inv;r-HO1-r-IFP-T; ptet-IFP
  • Ligation: J23119+r-CHE-T; ptet+r-CHE-T; ptet+r-HO1-r-IFP-T; J23119+Inv

28th July

  • PE
  • DD: r-HO1; r-GFP-T; J23119;r-CHE-T;r-HO1-r-IFP-t; ptet; r-CHE-T
  • Ligation: r-HO1 +r-GFP-T; J23119 +r-CHE-T; J23119 + r-HO1-r-IFP-T; pTet +r-CHE-T; pTet +r-GFP-T

30th July

  • PE
  • DD : J23119-r-CHE-T; ptet-r-CHE-T; J23119-r-HO1-r-IFP-T; ptet-r-HO1-r-IFP-T; r-HO1-r-GFP-T; J23119; pTet; TetR; rbs
  • Ligation: r-HO1 +r-GFP-T; J23119 +r-CHE-T; J23119+r-HO1-IFP-T; ptet+r-CHE-T; ptet+r-GFP-T


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