Protocol tips
| Upstream tips |
Protocol tips |
Downstream tips |
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0.5 µg PCR product (purified with the Illumina GFX PCR DNA and gel band purification kit, Amersham, according manufactureŕs instructions) were digested with 5 U SfcI (NEB) in 50 µl reaction, according manufactureŕs instructions. To reach the complete digestion, after 1.5 h incubation at 37°C, 5 U SfcI more were added to each reaction and the incubation was prolonged overnight at 30°C. |
Reaction products were analyzed on an 8% acrylamide gel with ethidium bromide staining. |
| Protocol tips |
| 0.5 µg PCR product (purified with the Illumina GFX PCR DNA and gel band purification kit, Amersham, according manufactureŕs instructions) were digested with 5 U SfcI (NEB) in 50 µl reaction, according manufactureŕs instructions. To reach the complete digestion, after 1.5 h incubation at 37°C, 5 U SfcI more were added to each reaction and the incubation was prolonged overnight at 30°C. |
| Downstream tips |
| Reaction products were analyzed on an 8% acrylamide gel with ethidium bromide staining. |
Publication protocol
0.5 µg PCR product (purified with the Illumina GFX PCR DNA and gel band purification kit, Amersham, according manufactureŕs instructions) were digested with 5 U SfcI (NEB) in 50 µl reaction, according manufactureŕs instructions. To reach the complete digestion, after 1.5 h incubation at 37°C, 5 U SfcI more were added to each reaction and the incubation was prolonged overnight at 30°C. Reaction products were analyzed on an 8% acrylamide gel with ethidium bromide staining.
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