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Let's make a primer! DNA to be amplified:. Use nearest-neighbour algorithm (recommended):. This is freaking awesooooooome. Punch in your 5'-3' DNA segment that you want amplified and hit the button with the dirty word. Optimal melting temperature for the primer. Generally, Primer Maker will come up with primers 1 to 2 degrees from the target temp. Concentration of Na and Mg in the buffer. The default values work well enough for the NEB buffers. Uses an awesome algorithm to calculate Tm. Don't turn th...
