10.3.2.4. Ligation of MLPA half-probes


1. Reduce the temperature of the thermal cycler to 54°C.  
2. While at 54°C, add to each sample:
2.1.  3µl                  Ligase-65 buffer A,
2.2.  3µl                  Ligase-65 buffer B,
2.3.  25 µl               sterile water,  
2.4.  1 µl                 Ligase-65.
3. Mix well.
4.  Incubate:
  4.1.  10-15 min at 54°C,
  4.2.   5 min at 98°C (inactivation of Ligase-65).
5. Cool on ice.