MidRange II PFG Marker

Description

Properties and Usage

Bases

Effective Size Range

Storage Temperature

Storage Conditions

10 mM Tris-HCl
1 mM EDTA
50% Glycerol
1% LMP agarose
pH 8.0 @ 25°C

Notes

1. The photograph represents the pulsed field gel separation of Mid Range II Markers 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.
2. 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. Allow no bubbles to form.
3. Recommended plug sizes are from 5-10 µl. A 10 µl plug (one small graduation on the GelSyringe volume scale) contains approximately 0.5 µg of DNA. Each GelSyringe yields 50+ plugs.
4. Melting plugs will cause denaturation of concatemers.
5. Place plug at the front of the well and seal with molten agarose just above gelling temperature(~42–45°C). Allow no bubbles to form. 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.

References

1. Greci, J. and Davis, T.B., New England Biolabs, unpublished observation.

Supporting Documents

Material Safety Datasheets

• MidRange II PFG Marker MSDS