Bisulfite-Seq High Sensitivity Kit (For Illumina®) (ab185907)
Key features and details
- Assay time: 8 hr
- Sensitivity: 0.5 ng
Overview
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Product name
Bisulfite-Seq High Sensitivity Kit (For Illumina®) -
Sensitivity
Assay time
8h 00mProduct overview
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.
Notes
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.
Properties
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Storage instructions
Please refer to protocols. -
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 -
Research areas
Properties
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Storage instructions
Please refer to protocols. -
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 -
Research areas
Images
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Size distribution of library fragments
Size distribution of library fragments. Post-bisulfite DNA library was prepared from 10 ng of input DNA using ab185907.