TECHNICAL DATASHEETDNA methylation control package V2Cat. No. C02040019
Format: 48 rxns
Product contentComponent

Quantity

Concentration

Storage

Methylated spike-in control

70 µl

0.1 ng/μl-20°C/-4°F

Unmethylated spike-in control

70 µl

0.1 ng/μl-20°C/-4°F

Primer pair for methylated spike-in ctrl

144 µl

5 µM each-20°C/-4°F

Primer pair for unmethylated spike-in ctrl

144 µl

5 µM each-20°C/-4°F

Product description
The DNA methylation control package V2 includes one methylated and one unmethylated spike-in controls together with theircorresponding qPCR primer sets that can be added to the DNA sample of interest for any methylation profiling experiment.
Those spike-in controls have been produced using synthetic oligonucleotides and are not homologous to any model species.
They will not interfere with the DNA sample of interest.
Applications

For bisulfite sequencing (e.g. RRBS - Reduced Representation Bisulfite Sequencing)
The spike-in controls must be added to the DNA sample of interest before bisulfite conversion and library preparation to monitor
the efficiency of bisulfite conversion. Because their methylation status is known, they can be used to assess the conversion
rate, i.e. in the case where you sequence the fully methylated control, all reads should align only to the methylated reference
sequence if the conversion ratio is 100%. The sequences of these controls are shown in Table 1.

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Diagenode Inc. USA | NORTH AMERICA

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Last update: January 20, 2021

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TECHNICAL DATASHEET
Table 1. Sequences of the methylated and unmethylated spike-in controls
Spike-in control

StrandSequence (5’>3’)

ForwardTCGAACGCCCGGTCGCGCGTGTGGAGGATGTCGGTGCTGCAGGGGGCGTTGGAGCCGAAGTAGACCGCGCCGAACGTACGAACGGTCGTTCATATTAGTTATACTGGTTGTCTTGA
TGTTGCTTCTCCTAATTTReverse

AAATTAGGAGAAGCAACATCAAGACAACCAGTATAACTAATATGAACGACGACCGTTCGTACGTTCGGCGCGGTCTACTTCGGCTCCAACGCCCCCTGCAGCACCGACATCCTCCACACGCGCGACCGGGCGTTCGAForwardTmCGAGAGGmCAGTGGGTGmCTGTmCGGmCGAAGTAGGGGTTGAmCGATGmCmCGGAmCmC
GTTmCGTAmCGTAmCGmCGmCGAmCmCGmCTAmCTAATTTTAGT
TGTGTTTmCTGGTmCmCTATTGATmCATTATmCTTGC

ReverseGmCAAGATAATGATCAATAGGAmCmCAGAAACACAAmCTAAAATTAGTAGmCGGTmCG
mCGmCGTAmCGTAmCGAAmCGGTmCmCGGmCATmCGTCAAmCCCCTAmCCTTmCGCmCG
ACAGmCAmCmCmCAmCTGmCmCTmCTmCGA

Unmethylated

MethylatedFor more details on the analysis of those spike-in controls after bisulfite sequencing, please download our Premium RRBS spike-in controls
manual.

For capture of methylated DNA (e.g. MeDIP - Methylated DNA Immunoprecipitation)
The spike-in controls must be added to the DNA sample of interest before the capture. The package also includes primer pairs
targeting respectively the methylated and unmethylated spike-in controls, allowing to check the success of the capture by
qPCR. The size of the expected amplicons is described in Table 2.
Table 2. Expected PCR product size
RegionAmplicon size

Unmethylated spike-in control

110 bp

Methylated spike-in control

103 bpStorage conditions: Store at -20°C/-4°F. For long storage, store at -80°C /-112°F. Avoid multiple freeze-thaw cycles.
Precautions: This product is for research use only. Not for use in diagnostic or therapeutic procedures.

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