Colony PCR protocol

1. Grow fresh colonies from transformation or single colony streaks overnight on LB selective media.
2. Using a sterile plastic pipette tip (20 or 200 µl), pick at least ¼ of a single, isolated colony.
3. In a PCR tube, resuspend cells in dH₂0 at room temperature and 1X primers to 25 µl total. Add 25 µl of Monserate Supernova hot start Taq mix, and place in PCR machine.
4. Amplify with primer/template specific conditions.

Example recipe and cycling parameters:

5 µl right primer (2 uM)
5 µl left primer (2 uM)
15 µl dH20 + cells
25 µl 2X SuperNova PCR mix.
50 µl total

Sample Thermocycler program:

1-10 min 95°C for Taq activation and bacterial lysis
2-95 7°C, 30 seconds
3-57°C, 30 seconds
4-72°C, 1 minute (for a 392 bp PCR product)
5-Repeat steps 2-4 29 times (30 cycles total)

Have a protocol or tip for using Monserate products you’d like to share? Send us a note:info@monseratebiotech.com