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ab185906 is a complete set of optimized reagents to prepare a DNA library -- after successful bisulfite conversion -- for various Illumina® platform-based bisulfite sequencing (bisulfite-seq) assays, such as whole genome bisulfite sequencing (WGBS), oxidative bisulfite sequencing (oxBs-seq), reduced representative bisulfite sequencing (RRBS), and other bisulfite-based next generation sequencing applications. The optimized protocol and components of the kit allow both non-barcoded (singleplexed) and barcoded (multiplexed) DNA libraries to be quickly constructed using sub-nanogram input concentrations of DNA since the DNA is first bisulfite-converted and then used for library preparation.
Illumina® is a registered trademark of Illumina, Inc.
Several methods, including whole genome bisulfite sequencing (WGBS) and reduced representation bisulfite sequencing (RRBS), are currently used for genome-wide DNA methlyation analysis. These methods convert unmethylated cytosines to uracil while 5-methylcytosines remain unchanged by the bisulfite treatment. This allows epigenetic differences to become genetic differences, which can be subsequently detected via sequencing at the single-based resolution level and on a genome-wide scale. However, practical use with such current methods are not ideal as they (1) need large amounts of DNA (>1 µg) as input material, which is difficult to prepare from limited biological samples such as tumor biopsy, early embyros, embyronic tissues, and circulating DNA; (2) require DNA to first be sheared and then ligated to adapters, followed by bisulfite conversion (post-ligation bisulfite conversion), which causes substantial amounts of DNA fragments contained in the adapter-DNA fragment constructs to be broken and thereby forms mono-tagged templates that become removed during library enrichment; and (3) are time-consuming (2 days). The Post-Bisulfite DNA Library Preparation Kit (For Illumina®) is designed to overcome these weaknesses.
|Components||12 tests||24 tests|
|10X dA-Tailing Buffer||1 x 40µl||1 x 80µl|
|10X End Repair Buffer||1 x 40µl||1 x 80µl|
|2X HiFi PCR Master Mix||1 x 160µl||1 x 320µl|
|2X Ligation Buffer||1 x 250µl||1 x 500µl|
|5X Conversion Buffer||1 x 50µl||1 x 100µl|
|Adaptors (50 μM)||1 x 15µl||1 x 30µl|
|Conversion Enzyme Mix||1 x 15µl||1 x 30µl|
|Conversion Primer||1 x 26µl||1 x 52µl|
|Elution Buffer||1 x 1ml||1 x 2ml|
|End Repair Enzyme Mix||1 x 25µl||1 x 50µl|
|Klenow Fragment (3’-5’ exo-)||1 x 15µl||1 x 30µl|
|MQ Binding Beads||1 x 1.8ml||1 x 3.6ml|
|Primer I (10 μM)||1 x 15µl||1 x 30µl|
|Primer U (10 μM)||1 x 15µl||1 x 30µl|
|T4 DNA Ligase||1 x 15µl||1 x 30µl|
ab185906 has not yet been referenced specifically in any publications.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"