Difference between revisions of "DNA Handling"
Jump to navigation
Jump to search
(Created page with " 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. 1. Quick spin lyophilized DNA in a micro...") |
|||
| Line 1: | Line 1: | ||
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. | Use 1uL of the DNA when cloning. | ||
Latest revision as of 05:52, 30 August 2014
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.