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==== Things to do in order ==== * Set up capture reaction in total 25 μL in 8-tube strip (96-well plate if more samples are processed) with the following components: 400 ng of WGBS DNA, 200:1 of probe to target ratio and 1x Ampligase Buffer. # Calculate volume of WGBS libraries required for 400ng ## Adjust total volume to 50ul (could be any number) ## Evaporate WGBS libraries in 1.5mL LoBind tube to reduce total volume to fit 25ul capture reaction including probes and 10X Ampligase Buffer ## Resuspend evaporated WGBS libraries with H2O and transfer to 0.2mL 8-tube strip or 96-well plate based on number of sample # Calculate the amount of each probe set required. The amount is based on probe to target ratio and template amount. # Prepare probes and 10X Ampligase buffer mix ## Add probe/buffer mix to the tube with WGBS libraries ## Mix well (spinand layer with equal volume of mineral oil (25ul of mineral oil to 25ul of capture reaction) ## Spin down the tube again before incubate on thermocycler as the following program: 94 ºC for 30 s, gradually lower temperature to 55 ºC at 0.02 ºC/s in a thermocycler, and let hybridize at 55 ºC for 20 hours (This is requirement. Not recommend to incubate shorter than 20h)
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