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ab185907 is designed to carry out bisulfite conversion, followed by a "post-bisulfite" library preparation process for Illumina® platform-based bisulfite sequencing, all in one kit. The DNA library is constructed directly from bisulfite-converted DNA generated from a small amount of input DNA (500 pg to 500 ng). Intended applications include whole genome bisulfite sequencing, oxidative bisulfite sequencing, reduced representative bisulfite sequencing, and various other bisulfite-next generation sequencing techniques. The optimized protocol and components of the kit allow the DNA to be bisulfite converted and fragmented simultaneously followed by quick non-barcoded (singleplexed) and barcoded (multiplexed) library construction using sub-nanogram quantities of bisulfite converted DNA.
DNA methylation occurs by the covalent addition of a methyl group (CH3) at the 5-carbon of the cytosine ring, resulting in 5-methylcytosine (5-mC). DNA methylation is essential in regulating gene expression in nearly all biological processes including development, growth, and differentiation. Alterations in DNA methylation have been demonstrated to cause a change in gene expression. Genome-wide analysis of DNA methylation could provide valuable information for discovering epigenetic markers used for disease diagnosis, and potential targets used for therapeutics. Bisulfite sequencing via next-generation sequencing technologies allow for high volume, lower cost output of DNA sequence data towards a better understanding of DNA methylation.
|Components||24 tests||12 tests|
|10X dA-Tailing Buffer||1 x 80µl||1 x 40µl|
|10X End Repair Buffer||1 x 80µl||1 x 40µl|
|2X HiFi PCR Master Mix||1 x 320µl||1 x 160µl|
|2X Ligation Buffer||1 x 500µl||1 x 250µl|
|5X Conversion Buffer||1 x 100µl||1 x 50µl|
|Adaptors (50 μM)||1 x 30µl||1 x 15µl|
|Conversion Enzyme Mix||1 x 30µl||1 x 15µl|
|Conversion Primer||1 x 52µl||1 x 26µl|
|Desulphonation Solution||1 x 140µl||1 x 70µl|
|DNA Binding Solution||1 x 12ml||1 x 6ml|
|Elution Buffer||1 x 2ml||1 x 1ml|
|Elution Solution||1 x 1ml||1 x 0.5ml|
|End Repair Enzyme Mix||1 x 50µl||1 x 25µl|
|F-Collection Tube||1 x 30 units||1 x 15 units|
|F-Spin Column||1 x 30 units||1 x 15 units|
|Klenow Fragment (3’-5’ exo-)||1 x 30µl||1 x 15µl|
|Modification Buffer||1 x 6ml||1 x 3ml|
|Modification Powder||1 x 4 vials||1 x 2 vials|
|MQ Binding Beads||1 x 3.6ml||1 x 1.8ml|
|Primer I (10 μM)||1 x 30µl||1 x 15µl|
|Primer U (10 μM)||1 x 30µl||1 x 15µl|
|T4 DNA Ligase||1 x 30µl||1 x 15µl|
Size distribution of library fragments. Post-bisulfite DNA library was prepared from 10 ng of input DNA using ab185907.
ab185907 has not yet been referenced specifically in any publications.