User:Anhthai

Monday 22 January 2018

Prepare electrodes

• 80% Ag and 20% graphene.
• Firstly, wash the mixing vial with ethanol then acetone carefully to remove the remained Silver.
• Mix 4g Ag with 1g graphene homogeneously (before doing that, use 2 Al foil, 1 foil taking graphene to dry acetone and then take 1g graphene).
• Add 4uL acetone (after checking viscosity of the mixing solution).

Wednesday 24 January 2018

Test HPV-16

• There are two wells, 1st well for adding HPV-16 and 2nd well for adding control (primers).
• Attach and first measure the chip. (1st measurement)
• Each well, do the same protocol: 4uL then dry, 4uL then dry, 2uL then dry completely.
• Add 4uL DI water (well 1), wait 30sec and then measure impedance (2nd measurement).
• Add 4uL DI water (well 2), wait 30sec and then measure impedance (3rd measurement).

• Result: 64MOhm, 1.67MOhm, 1.55MOhm
• Comment: result fits with concept.

LAMP experiment

• Continue as yesterday we delayed.
• Check protocol to make HPV-16 LAMP.

Monday 29 January 2018

HCV primers literature review

Primer Length Genome position Sequences F3 19 88–106 5’ -TCCCGGGAGAGCCATAGTG-3’ B3 21 254–274 5’ -CACTCGCAAGCACCCTATCAG-3’ FIP 43 148–166 108–127 5’ -GATCCAAGAAAGGACCCGGTTTTTCTGCGGAACCGGTGAGTAC-3’ BIP 43 179–197 232–25 5’ -CCTGGAGATTTGGGCGTGCTTTTAGTACCACAAGGCCTTTCGC-3’ LF 17 129–145 5’ -TCATCCTGGCAATTCCG-3’ LB 18 202–222 5' -GCGAGACTGCTAGCCGAG-3’

Wednesday 31 January 2018

Answer question: What is available commercialized enzyme that can digest double stranded DNA at high temperature (like 80 C degree?)

• Search on NEB website:
  • ApeKI: Incubate at 75°C, no heat inactivation.

Thursday 1 February 2018

Sensitivity test with HPV sample

Monday 26 Mar 2018

HCMV Bubble project

PMMA chip fabrication

• two channels chip
• Setting:
  • Red 40 40 50
  • Black 80 2.3 100
• Adjust height for focusing
• Clean white powder on the top of the chip with ethanol.
• Attach DSA on the chip, after that we had to clean with H2O2 (avoid the buuble during adhesion process).

Nano probe

• 3mL of Pt NPs in glass bottle with magnetic beads, stirring at 110rpm. Cover in aluminium foil.
• Add drop-wise 500uL of Ab and PDPH.
• Incubate for 3 hours, room temperature, in dark at 110rpm.
• Concentration the solution using ultra-filltration 3000rpm at 10 mins.
• The remaining volume should be 300uL.

Antibody oxidation

• the stock is adjusted to 6.4 mg/mL
• 3.5uL of Ab + 96.5uL sodium acetate (at 0.1M pH 5.5)
• 100uL sodium metaperiodate (10mM), the total volume is now 200uL.
• 20 mins on ice and dark in Al foli.
• Add 1.5 times volume of PB at pH 7.4 300uL.
• total 500uL and the final concentration is 90ug/mL
• Add 500uL of 90ug/uL PDPH.
• Mix, incubate at room temperature, dark and total volume 1mL.
• Final concentration of Ab is 45ug/uL

Major steps in HCMC bubble projects

• Attach PMMA and DSA, clean slides with ethanol and silane.
• Proceed with oxygen plasma and silanization.