Thermo Scientific™ CpG Methylated Jurkat Genomic DNA

Genomic DNA was isolated from Jurkat E6.1 (human acute T-cell leukemia, clone E6.1) cells grown to confluency in RPMI media supplemented with 10% bovine fetal serum. Isolated genomic DNA was treated with CpG Methyltransferase, repurified and dissolved in TE buffer. ≥98% CpG methylation level confirmed by bisulfite sequencing.

Thermo Scientific CpG Methylated Jurkat Genomic DNA is human male genomic DNA from Jurkat cells, enzymatically methylated with CpG Methyltransferase (M.SssI). Genomic DNA was isolated from Jurkat (human acute T-cell leukemia) cells grown to confluency in RPMI media supplemented with 10% bovine fetal serum. Isolated genomic DNA was treated with CpG Methyltransferase, repurified, and dissolved in TE buffer. ≥98% CpG methylation level confirmed by bisulfite sequencing.

Highlights:
• Pure - DNA free from inhibitors, proteins, and RNA
• Convenient - ready to use 100 ng/μL concentration
• Compatible with downstream applications
• Complete methylation of cytosines in CpG dinucleotides

Applications
As controls in the following:
• Bisulfite sequencing
• Combined Bisulfite Restriction Analysis (COBRA)
• Methylation-sensitive Single-Nucleotide Primer Extension (Ms-SNuPE)
• A260/230 ratio: ≥ 2
• A260/280 ratio:1.75 to 1.98

Quality Control
• Predominant > 30 kb band is observed during agarose gel electrophoresis. DNA concentration is determined spectrophotometrically. The absence of nucleases is confirmed by labeled oligonucleotide tests.
• CpG methylated DNA was protected from digestion by the methylation sensitive restriction enzyme HpaII as determined by qPCR analysis. Following qPCR, equivalent Cq values for HpaII digested and control undigested DNA are observed.
• DNA purity is confirmed through a functional qPCR test in which Cq values of MspI and HpaII digested template DNA must differ by ≥ 5 cycles.
• The absence of mycoplasma DNA is confirmed by PCR.
• As controls in bisulfite sequencing, Combined Bisulfite Restriction Analysis (COBRA), Methylation-sensitive Single-Nucleotide Primer Extension (Ms-SNuPE)