Transformation

Preparation

• iGEM parts / ligation products
• LBbroth (No antibiotic)　500μL
• TE　15μL
• plates
• ice box
• heat block(42℃)
• competent cells

→ always onice! Melt on ice! Mix DNA as soon as cells melt!

Protocol

to thaw out igem parts

• 1. With a pipette tip, punch a hole in the foil
• 2. Add 15uL of TE (MilliQ),and pipetting
• 3. Pipette 1uL of the resuspended DNA Transformation into your desired competent cells
• 4. Hold on ice for 30 mins
• 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℃ for 30 mins
• 9. Plate out
• 10. Incubate at 37°C