MidRange II PFG Marker

  • This product was discontinued on 01/01/2013
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Discontinued Products

Description

MidRange Marker II is concatemers of λ DNA isolated from the bacteriophage λ (cI857 ind1 Sam7) mixed with XbaI digested λ DNA embedded in 1% LMP agarose and supplied in a GelSyringe™ dispenser. XbaI produces fragments of 24.0 and 24.5 kb. These fragments anneal to and form concatamers with intact λ DNA. It is designed for use as a size marker for pulsed field gel electrophoresis (PFG). Size range: 24-300 kb.



N3552_thumb

1% agarose gel, 6 V/cm, 15°C for 24 hours. Switch times ramped from 1-25 seconds.

Properties and Usage

Bases

Fragmentbp
18291000
17267000
16266500
15242500
14218500
13218000
12194000
11170000
10169500
9145500
8121500
7121000
697000
573000
472500
348500
224500
124000

Effective Size Range

24,000bp to 291,000bp

Storage Temperature

-20°C

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 Greci, J. and Davis, T.B., New England Biolabs, unpublished observation.

FAQs

  1. There are several factors that lead to poor results for PFG markers. Here is a list of recommendations for ensuring a sharp high quality banding pattern.

Interactive Tools

Safety Data Sheet

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.