Table 1 Reagents used for the DNA methylation and restriction enzyme cleavage
From: Systematic genome sequence differences among leaf cells within individual trees
Step 1. Methylation reaction (10 μl) from the protocol of New England Biolabs Inc. | |
Nuclease free water | 6 μl |
10× HpaII methyltransferase buffer | 1 μl |
SAM (80 μM) | 0.1 μl |
Genomic DNA (10 ng) | 0.4 μl |
HpaII methyltransferase | 2.5 μl |
Step 2. HpaII digestion reaction (50 μl) | |
Methylation product (taken from step 1) | 10 μl |
10× NEBuffer 1 | 4 μl |
MgCl2 (10 mM) | 20 μl |
HpaII restriction enzyme | 4 μl |
Nuclease free water | 12 μl |