KAPA Hyper Prep and HiFi Uracil+ Kits Allow for Reduced Representation Bisulfite Sequencing of Ultra-low Amounts of DNA in B Cells

17 Dec 2015

This application note provides a robust methodology for RRBS analysis using the streamlined chemistry of the KAPA Hyper Prep Kit and HiFi Uracil+ HotStart DNA Polymerase, and demonstrates its utility in accurately quantifying the DNA methylation status from sample-limiting conditions. Methyl-Seq is a powerful tool for understanding genome-wide methylation with base-pair resolution. Several strategies have been developed to understand the methylation status of a genome, including reduced representation bisulfite sequencing (RRBS). Bisulfite treatment results in a significant decrease in DNA input and quality; therefore, the construction of high-quality libraries and efficient downstream amplification is critical.

Related Products

Request Quote for All Products

KAPA HiFi Uracil+

Kapa Biosystems, Inc.

KAPA HiFi DNA Polymerase provides high fidelity next-generation sequencing library amplification with the highest yield, lowest bias, and most uniform sequence coverage available. These characteristics are retained in KAPA HiFi Uracil+ DNA Polymerase, a modified version of KAPA HiFi engineered to tolerate uracil residues. The new enzyme is particularly well suited for applications employing bisulfite DNA conversion, which typically produces low concentrations of AT-rich DNA. KAPA HiFi HotStart Uracil+ ReadyMix (2X) is a ready-to-use cocktail containing all components required for PCR, except primers and template. Features: Higher yields and reduced size bias More uniform sequence coverage Improved representation of AT-rich sequences Applications: Methylation analysis – amplification of bisulfite-converted DNA for sequencing Amplification of damaged DNA samples Prevention of false-positive results due to carryover amplicon contamination

(0)