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