mTet1CD was prepared as described previously (17 (link)). DNA was incubated in a 50-μl reaction containing 50 mM Hepes buffer (pH 8.0), 100 μM ammonium iron (II) sulfate, 1 mM α-ketoglutarate, 2 mM ascorbic acid, 2 mM dithiothreitol, 100 mM NaCl, 1.2 mM adenosine triphosphate, and 4 μM mTet1CD for 80 min at 37°C. After that, 0.8 U of proteinase K (New England Biolabs) was added to the reaction mixture and incubated for 1 hour at 50°C. The product was cleaned up on Bio-Spin P-30 Gel Column (Bio-Rad) and 1.8× AMPure XP beads following the manufacturer’s instruction.
Bio spin p 30 gel column
Manufactured by Bio-Rad
The Bio-Spin P-30 Gel Column is a size-exclusion chromatography column designed for the purification and desalting of biomolecules. It is used to separate molecules based on their size and molecular weight, allowing for the removal of small molecules, salts, or other contaminants from the sample.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using bio spin p 30 gel column
1
Methylation Profiling of DNA by mTet1CD
2
Enzymatic DNA Modification Protocol
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Genomic DNA (up to 200 ng) was incubated in a 50 μl reaction containing 50 mM HEPES buffer (pH 8.0), 100 μM ammonium iron(II) sulfate, 1 mM α-ketoglutarate, 2 mM ascorbic acid, 1 mM dithiothreitol, 100 mM NaCl, 1.2 mM ATP and 4 μM mTet1CD for 80 min at 37 °C. After that, 0.8 U of Proteinase K (New England Biolabs) were added to the reaction mixture and incubated for 1 h at 50°C. The product was cleaned up on Bio-Spin P-30 Gel Column (Bio-Rad) and 1.8× AMPure XP beads following the manufacturer's instruction.
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