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Our cloning strategy requires 20ng of Casein-DNA in 1uL. This means we need a final concentration of DNA for cloning to be 20ng/uL.
- Quick spin lyophilized DNA in a microcentrifuge.
- Add 10uL of 1x Low TE Buffer (Tris/EDTA- low refers to the concentration of EDTA. If EDTA is too high in your final cloning reaction, you can inhibit Cation-dependent enzymatic reactions. EDTA is a di-cation chelator and pretty much absorbs all free Ca2+, Mg+ and Mn+ in DNA-based reactions.
- Flick the tube - vigorously.
- Quick spin tube again (15s)
Use 1uL of the DNA when cloning.