Lab Notebook

January (Week 1-4)

Brainstorming and deciding the direction of the project.

February (Week-5)

  • Prior to any wet-lab experiments, the promoter sequences of ADA-regulon were analyzed and their binding sites were taken into consideration.

  • Ecocyc was used to extract the desired sequences.

  • Snapgene was used for the selection of compatible cloning vectors and their overhangs.

  • Primer designing was done with the help of NEB cutter V2.0, NEB Tm calculator and NEB: Double digest finder.

  • Primers were ordered.


  • PCR amplification of promoter of ADA-AlkB gene followed by gel electrophoresis.

  • PCR amplification of the reporter gene from EGFP-N1 plasmid followed by gel electrophoresis.

  • Gel extraction of amplified sequences.


  • Sequential restriction digestion of extracted Ada-AlkB promoter using HindIII and EcoRI.

  • Sequential restriction digestion of extracted reporter sequence using EcoRI and BamH1.


  • Ligation of the digested sequences was done using T4-DNA Ligase.

  • This ligated DNA was transformed into DH5-alpha E.coli competent cells followed by overnight incubation.

  • Colonies were selected for colony PCR.

March (Week-9)

  • The transformed colonies were checked again for the desired DNA using plasmid isolation. MDI Mini-prep kit was used for plasmid isolation.

  • This was followed by their restriction digestion.

  • Gel electrophoresis was conducted.


  • Gel images were analysed.

  • Orders were placed for analytes for conducting assay experiments by artificially spiking water samples.

  • Our institute was closed due to COVID 19 and the entire state was under lockdown.