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Team:UT-Tokyo/LabWork/RegularMethods
From 2012.igem.org
Regular Methods
Assembly parts
Digest
Materials
Protocol
Ligation
Materials
- Vector DNA
- Insert DNA
- 2x Ligation Mix
Protocol
1.Make reaction liquid
- MilliQ up to 20uL
- 10uL 2x Ligation Mix
- Vector DNA
- Insert DNA
2.Incubation at 16 °C for 15-30 min.
Transformation
Materials
- BioBrick parts / ligation products
- SOC or LB (No antibiotic) 500uL
- TE 15uL
- plates
- competent cells
Protocol
to thaw out igem parts
1.With a pipette tip, punch a hole in the foil
2.Add 15μL of TE (MilliQ),and pipetting
3.Pipette 1μL of the resuspended DNA Transformation into your desired competent cells
4.Hold on ice for 30 min.
5.Heat shock at 42°C for 45 seconds (and on ice after it)
6.Add 300uL of LBborth in each epp
7.Wait for 10 mins
8.Hold at 37°C for 30 min.
(this step can be skipped with ampicillin selection)
9.Plate out
10.Incubate at 37°C
Purification of DNA
Reagents
