Protocol tips
Upstream tips |
Protocol tips |
Downstream tips |
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For best results, the CT Conversion Reagent should be prepared freshly and used immediately following preparation. |
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Protocol tips |
For best results, the CT Conversion Reagent should be prepared freshly and used immediately following preparation. |
Publication protocol
Bisulfite sequencing
10 ng of genomic DNA was treated with sodium bisulfite to convert unmethylated cytosines to uracils with the EpiTech Bisulfite Kit according to the manufacturer’s protocol (Qiagen, Inc). Bisulfite converted DNA was amplified by PCR and the amplicons cloned using Topo TA vectors (Invitrogen). Positive colonies were purified and sequenced (Beckman Coulter Genomics). Bisulfite sequences were aligned to their respective in silico bisulfite converted genomic sequences using the Bioconductor Biostrings R package and custom R scripts, which are available upon request. Only sequences that contained data for all CpGs and with a bisulfite conversion rate of greater than 95% (as determined by conversion of non-CpG cytosines to uracil and subsequent amplification as thymine) were included in the analysis. Significance was determined by a one sided Fisher’s exact test.
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Papers
Check out relevant papers found by Labettor's AI that are relevant for performing DNA methylation profiling Whole genome profiling - mouse T-cell (CD4 / CD8) using EpiTect Bisulfite Kit from Qiagen.
Paper title
Global DNA methylation remodeling accompanies CD8 T cell effector function
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