User:Jarle Pahr/Protocols
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Links to various protocols:
Mixing of dNTPs.
A 10 mM dNTP mix should contain 10 mM dNTPs total (2.5 mM of each). To prepare 40 uL 10 mM dNTP, mix:
1 uL 100 mM dATP 1 uL 100 mM dGTP 1 uL 100 mM dCTP 1 uL 100 mM dTTP 36 uL H2O.
Ligation:
~100 ng equivalents backbone DNA. 3:1 - 5:1 molar ratio insert/backbone 2 uL T4 DNA ligase buffer 0.5 uL T4 DNA ligase H2O to 20 uL
Incubate at 16 C for 3 h or overnight.
