Find your Plasmid Pronto with SapphireAmp® Fast PCR Master Mix
Colony PCR is a method used to screen for plasmids containing a desired insert directly from bacterial colonies without the need for bacterial culturing or plasmid purification steps. SapphireAmp Fast PCR Master Mix further streamlines the colony PCR workflow by allowing PCR reactions to be assembled by simply adding primers and cells to the premix. After PCR, the reaction mixture can be loaded directly onto a gel for electrophoresis or used directly for restriction enzyme digestion of PCR products. SapphireAmp master mix will streamline and speed up your workflow, allowing screening of inserts of up to 5 kb in only 1.5 hours.
Streamlined Colony PCR Protocol using SapphireAmp Fast PCR Master Mix
"I used SapphireAmp for colony PCR. It works super fast and the result is reliable. I like the fact that the dye is already included in the master mix." -Researcher from Rhode Island Hospital
Using SapphireAmp Fast PCR Master Mix to Screen a cDNA Library by Colony PCR
E. coli cells were transformed with a mouse cDNA library (insert size from 0.5 to 5 kb) ligated into pUC118. Plasmid-containing transformants were analyzed using SapphireAmp Fast PCR Master Mix and M4 and RV primers. The PCR reaction cycle was complete in just 70 minutes, and excellent yields were obtained. All 16 clones checked were confirmed to contain 0.5 to 5 kb inserts. For additional experimental details, click here to download the entire application note.
Speed up Colony PCR with SapphireAmp Fast PCR Master Mix
SapphireAmp Fast PCR Master Mix is fast— with an extension speed of 10 seconds/kb, reactions can be completed in half the time of conventional Taq. SapphireAmp master mix is also significantly faster than dye-added premixes offered by other manufacturers. The table below lists the reaction time for SapphireAmp master mix and the recommended reaction times for two other commercially available dye-added premixes.