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Team:UT-Tokyo/LabWork/RegularMethods
From 2012.igem.org
(Difference between revisions)
| Line 31: | Line 31: | ||
*Insert DNA | *Insert DNA | ||
=====2.Incubation at 16 °C for 15-30 min.===== | =====2.Incubation at 16 °C for 15-30 min.===== | ||
| + | |||
| + | |||
==Transformation== | ==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== | ==Purification of DNA== | ||
Revision as of 21:50, 26 September 2012
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
