| Library |
| Library name |
Construction protocol |
Strategy |
Source |
Selection |
Layout |
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Genomic DNA of sorted pZscan4c-Em+, pZscan4c-Em- cells at stage II D12 and pZscan4c-Emerald XEN-like cell were extracted using EasyPure Genomic DNA Kit (TransGen, EE101-11). DNA were denatured and bisulfite converted using EZ DNA Methylation-GoldTM Kit (Zymo Research, D5005). In brief, 20 µl DNA (300 ng) was mixed with 130 µl CT Conversion Reagent and performming the following steps: 98°C for 10 minutes, 64°C for 2.5 hours. Samples and 600 µl M-Binding Buffer were added into a Zymo-Spin™ IC Column, followed by inverting the column several times and centrifuging at full speed (>10,000 x g) for 30 seconds. 100 µl M-Wash Buffer was added to the column and centrifuged at full speed for 30 seconds. Add 200 µl M-Desulphonation Buffer to the column, incubate at room temperature (20-30°C) for 15-20 minutes and then centrifuging at full speed for 30 seconds. Wash the column with 200 µl M-Wash Buffer twice and elute the DNA with 10 µl M-Elution Buffer. Recovered bisulfite-converted DNAs were constructed into sequencing libraries using Accel-NGS Methyl-Seq DNA Library Kit (Swift, 30024) following the manufacturer’s instructions. |
Bisulfite-Seq |
GENOMIC |
PCR |
PAIRED
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