Description:

MidRange PFG Marker consists of concatemers of λ DNA isolated from the bacteriophage λ (cI857 ind1 Sam7) mixed with XhoI digested λ DNA embedded in 1% LMP agarose and supplied in a GelSyringe™ dispenser. XhoI produces fragments of 15.0 and 33.5 kb. These fragments anneal to and form concatemers with intact λ DNA. It is designed for use as a size marker for pulsed field gel electrophoresis (PFG). Size range: 15-300 kb.1% agarose gel, 6 V/cm, 15°C for 24 hours. Switch times ramped from 1-25 seconds.

Notes:

The photograph represents the pulsed field gel separation of Mid Range PFG Marker using a CHEF apparatus. The 1% agarose gel was run at 6 volts/cm using ramped pulse times from 1 to 25 seconds for 24 hours at 15°C in 0.5X TBE (50 mM Tris-HCl, 50 mM boric acid, 1 mM EDTA) made with Milli-Q™ water.Extrude agarose from GelSyringe carefully and slice plugs from the end with a sharp blade. One plug is sufficient for one lane of a gel. Place the plug at the front of the well and seal with molten agarose just above gelling temperature (~42–45°C). Allow no bubbles to form.Recommended plug sizes are from 5-10 µl. A 10 µl plug (one small graduation on the GelSyringe volume scale) contains approximately 0.5µg of DNA. Each GelSyringe yields 50+ plugs.Melting plugs will cause denaturation of concatemers.Never attach the agarose plugs to the gel comb before the gel is poured. Heat from the solidifying gel will cause the Lambda concatemers to denature.