Detection of bisulfite modified DNA can be facilitated through conventional PCR, methylation specific PCR, and bisulfite genomic sequencing after PCR amplification with or without cloning. Want to learn more about the DNA methylation analysis workflow? Visit the Epigenetics Learning Center.

High Fidelity Results from Conventional PCR and Methylation Specific PCR* (MSP)

Following bisulfite conversion, the modified DNA template is distinguishable from the original template at methylated cytosines. This results in a pool of DNA fragments with altered nucleotide sequences due to differential methylation status. The methylation specific PCR reaction involves amplifying the altered nucleotide sequences using methylated specific primers for the CpG islands of interest.

Invitrogen’s PCR products are tailored to address the persistent challenges associated with amplifying bisulfite modified DNA - such as a small initial sample of highly fragmented DNA. Using the Platinum® PCR kits, you can expect:
  • Faithful amplification based on reliable and consistent single-nucleotide discrimination
  • High sensitivity and high-yield of bisulfite converted DNA
  • Room temperature reaction assembly

Simplify Your Preparation for Bisulfite Genomic Sequencing

Combine effective plasmid DNA purification with quick and efficient cloning. The PureLink™ Quick Plasmid Miniprep Kit provides efficient and reliable purification of plasmid DNA for cloning with TOPO® Technology. Click to learn more about additional plasmid DNA purification solutions.

TOPO® Technology presents a faster, more efficient way to clone PCR products for sequencing. The key to TOPO® Cloning is the enzyme DNA topoisomerase I, which functions as both a restriction enzyme and a ligase. Its biological role is to cleave and rejoin DNA during replication. To harness the religating activity of topoisomerase, over 30 vectors are provided linearized with topoisomerase I covalently bound to each 3' phosphate. This enables fast ligation of DNA sequences with compatible ends. After only 5 minutes at room temperature, the ligation is complete and ready for transformation into E. coli. 

Top-rated sequencing vectors

The TOPO® Cloning Kits for Sequencing contain vectors with a minimized multiple cloning site that positions the T7 and T3 priming sites only 33 bp away from the PCR product insertion site. This allows you to sequence more of your insert and less of the vector, saving you time and money. There are two vectors available:

  • TOPO TA Cloning® Kits for Sequencing – fast and efficient results from Taq-amplified PCR products
  • Zero Blunt® TOPO® PCR Cloning Kits for Sequencing – easiest and most effective method for  use with blunt-end fragments

Diagram Methylcode ERa
Figure 1. Conversion of unmethylated cytosines to uracil with bisulfite treatment

Catalog and Ordering

Get results using the Platinum “hot start” line of PCR products. Refer to the PCR enzyme selection guide for details and other PCR products.

Looking for primer design software for help with low complexity DNA? Use Invitrogen’s primer design tools.

* NOTE: Methylation specific PCR may be covered by one or more of U.S. Patents Nos. 5,786,146, 6,017,704, 6,200,756 and 6,265,171 and patents based upon foreign counter-part applications.  No license or rights under these patents to perform methylation specific PCR is conveyed expressly or by implication to the purchaser of this product. User’s of Invitrogen Corporation’s products subject to this label license should determine whether they have all the appropriate licenses in place. Further, no warranty is provided that the use of these products will not infringe on the patents referred to above.

Epigenetics Learning Center

Our Learning Center provides an overview of epigenetic regulation of gene expression as well as considerations underpinning the research workflow. Learn about:

microRNAs

DNA methylation

Support