Table 1.

PCR conditions for a dSLAM query construct.

Ingredient	Volume
10 x ExTaq (or LA Taq) Buffer	10 μl
dNTP (2.5 μM each)	8 μl
Forward primer (20 μM)	5 μl
Reverse primer (20 μM)	5 μl
Template DNA (200 ng/μl)	2 μl
ExTaq or LA Taq DNA polymerase (5 units/μl)	0.5 μl
dH2O	69.5 μl
Total	100 μl
94°C 5′; 35 x (94°C 20″, 55°C 20″, 72°C 3′30″); 72°C 5′; 4°C hold

Note: The PCR annealing temperature should be adjusted according to the melting temperatures (Tm) of the primers. We typically use primers with Tm values ranging from 52°C to 60°C. 5 μl of DMSO should be added to the PCR mixture if the template, for example the natMX cassette, is G/C-rich.