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Contents & Storage

Hydroxymethyl Collector™ contains all thre reagents needed to perform the glucosylation reaction, biotin conjugation, streptavidin capture, elution and purification of DNA containing 5-hydroxymethylcytosine. Protocols are provided for the fragmentation of sample DNA all the way through to real time PCR analysis of a locus of interest. The enriched dsDNA is compatible with Next-Generation sequencing library preparation. For added convenience, the kit also includes positive control 5-hmC DNA, which is a 338 bp sequence of the APC locus in which 25% of the cytosine residues have been hydroxymethylated. PCR primers for APC are provided to confirm the efficiency of the enrichment reactions.

Please note that the Hydroxymethyl Collector™ Kits are shipped on dry ice and contain reagents with multiple storage temperatures inside. Please store each component at the temperature indicated below. All reagents are guaranteed stable for 6 months from date of receipt when stored properly. Do not refreeze the streptavidin beads. Once thawed, store at their proper temperature. This kit includes the following components:

  • β-glucosyltransferase enzyme (200 U/µl); Store at -20°C
  • UDP-Azide-Glucose (3 mM); Store at -20°C
  • 10X Reaction Buffer AM1; Store at -20°C
  • 1 M DTT; Store at -20°C
  • Biotin Conjugate Solution; Store at -20°C
  • 5-hmC Control DNA (50 ng/µl); Store at -20°C
  • APC PCR Primer Mix (2.5 µM); Store at -20°C
  • Human genomic DNA, MseI digested (20 ng/µl); Store at -20°C (only in Hydroxymethyl Collector Catalog No. 55013)
  • Streptavidin Beads; Store at 4°C
  • Binding Buffer AM13; Store at 4°C
  • 10X Elution Buffer AM2; Store at 4°C
  • DNA Purification Binding Buffer; Store at RT
  • DNA Purification Wash Buffer; Store at RT
  • DNA Purification Elution Buffer; Store at RT
  • ChIP Filtration Columns; Store at RT
  • 3M Sodium Acetate; Store at RT
  • DNA purification columns & collection tubes; Store at RT
  • Bar magnet & glue dots; Store at RT