Noi/NOTES/2014-3-29
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DNA methylation sensitivity test assay - MONOD Project, MONOD_GP1_V4 probe set (BSPP capture)
- Continued from: [[1]]
- File:MONOD-sensitivity-test-plans.png
- - For all bis-DNA input, I will use the same concentration of probe in capture reaction by using 500:1 probe:target ratio for 300ng bis-DNA
BSPP capture set up
Probe calculation
Probe:target | 500:1 | Unit | Calculation |
Probe size (number of individual probe in oligo mix) | 7,110 | probes | |
Human bis-cvt gDNA required | 300 | ng | |
Human gDNA MW | 1.822E+12 | g/mole | 3.0E+09 * 607.4Da/bp |
Human gDNA( 300ng) | 1.646E-19 | mole | 300E-09g / 1.822E+12g/mole |
Probe required (500:1) | 8.232E-17 | mole | 500 * 1.098E-19mole |
Probe mix MW (size 7,110probes, 110nt) | 2.381E+08 | g/mole | 7,110 probes * (110bp *303.7Da/bp+79Da) |
Mass of probe required | 1.960E-08 | g | 8.232E-17mole * 2.381E+08g/mole |
Mass of probe required | 19.60 | ng | |
Probe conc | 6.87 | ng/ul | |
Volume of probe | 2.85 | ul |