QIAprep Spin Minprep Kit
- Pellet 1 - 5 mL bacterial overnight culture by centrifugation at >8000 rpm for 3 min at room temperature
- Resuspend pelleted bacterial cells in 250 µl Buffer P1 and transfer to microcentrifuge tube
- Add 250 µl Buffer P2 and mix by inverting tube 6 times until solution is clear.
- Add 350 µl Buffer N3 and mix by inverting the tube 6 times.
- Centrifuge for 10 minutes at 13,000 rpm
- Apply the supernatant liquid to the QIAprep spin by decanting. Centrifuge for 30-60 second and discard the flow-through.
- Wash the QIAprep column by adding .75 ml Buffer PE
- Centrifuge for 60 seconds and discard the flow through.
- Centrifuge for 1 min to remove residual wash buffer.
- Place the QIAprep column in a clean 1.5 ml tube. To remove the DNA, add water to the center of the QIA spin columns, and centrifuge for 2 min.
Transformation of Linker Genes
- Transformations: Linker genes from Genscript (4 total)
> Synthesized DNA from Genscript (red screw-cap tubes)
- PL rigid
- PL rigid 4
- PL flex
- PL flex 4
Note: these parts arrived in the pUC57 vector: 
> Warm five 100 μg/mL Amp agar plates at 37 °C
> Thaw fresh tube of DH5α Turbo cells on ice
> Resuspend invisible DNA pellet (stock DNA) in 10 μL dH2O
> Add 0.5 μL stock DNA to 10 μL dH2O in 0.5 mL tubes
> Add 30 μL of DH5α Turbo cells to DNA + dH2O
--> Include #5 tube, water only, no DNA (negative control)
> Incubate cells + DNA on ice for 5 min.
> Label pre-warmed plates
> Transfer cells + DNA onto agar
> Add 10 - 15 sterile glass beads, shake, discard beads
> Incubate plates at 37 °C overnight