Notebook – Protocol – 5. Restriction enzymes digestion

1. Restriction enzymes digestion was done using Fast Digest product (Thermoscientific) using the following composition:

• DNA sample (1-3 ug), 15 ul
• Fast Digest restriction enzyme #1, 2 ul
• Fast Digest restriction enzyme #2, 2 ul
• 10x buffer, 3 ul
• Sterilized distilled water, 6 ul
Total volume, 30 ul

2. Samples were incubated at 37 °C for more than 1 hour (typically 3 hours).

3. When necessary, samples were electrophoresed using protocol “3. Electrophoresis”(typically 3 hours).

4. Samples were purified using protocol “4. DNA purification”.

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