Table 1 Hybridization reaction conditions

From: Single-molecule force spectroscopy of toehold-mediated strand displacement

STEP

TEMP

TIME

First hybridization

 Initial Denaturation

70 °C

1 min

 Folding

68 °C

65 °C

30 s

20 min (30 cycles)

 Final

25 °C

5 min

 Hold

4–10 °C

∞

Second hybridization

 Initial Denaturation

68 °C

1 min

 Folding

68 °C

63 °C

30 s

20 min (30 cycles)

 Final

25 °C

5 min

 Hold

4–10 °C

∞

  1. In the first hybridization reaction, the toehold hairpin is annealed to a DNA handle and the adapter strand to another DNA handle. In the second hybridization reaction, the products of the first hybridization reaction are annealed together.
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