Workpackage 4: Nanopore Sequencing

Project 1: Exonuclease-Nanopore based Sequencing

1. 'Exonuclease selection and gene construction'
   Bioinformatic selection of exonucleases.
2. 'Exonuclease-nanopore fusion construction'
   Preparation of exonuclease-nanopore fusion constructs.
3. 'Addition of base discrimination to exonuclease-nanopore'
   Inclusion of base discrimination to nanopores.
4. 'Nanofluidic control of base from cleavage to site of detection'
   System for joining exonuclease and nanopore.

Project 2: Integration of Protein pores in solid-state apertures

1. 'Solid-state aperture formation'
   Preparation of arrayed apertures for nanopores.
2. 'Engineered α-HL protein pores with targeted delivery to apertures'
   Determination of conditions for good interaction of arrayed apertures and α-HL proteins.
3. 'Bonding of protein pores to aperture surface'
   Method for bonding α-HL to arrayed apertures.
4. 'Protein pores in solid state aperture arrays'
   Validation of arrayed apertures for nanopores.

Project 3: Droplet-based bilayers

1. 'Integration of DNA amplification with nanopore sensing'
   Investigation of nucleic acid amplification technologies with nanopore droplet bilayers.
2. 'Construction of electronic system and microfluidic devices'
   Preparation of a device for droplet-based bilayer nucleic acid amplification procedures.
3. 'In situ amplification and detection of SNPs'
   Validation of device of task 3.2.
4. 'Multiplexed system for haplotyping'
   Validation of device of task 3.2 for haplotyping.

Project 4: Methylometer

1. 'Base discrimination by protein nanopores'
   Detection of 5-methyl cytosine in a nanopore.
2. 'Decreased DNA translocation to improve base resolution'
   Decrease of nucleic acid translocation through a nanopore by the application of low affinity antibodies.
3. 'Double-stranded DNA epigenetic analysis'
   Investigation of double stranded nucleic acid translocation through nanopores.