For life science research only. Not for use in diagnostic procedures. DNA Molecular Weight Marker II (0.12–23.1 kbp) ␭DNA • Hind III digested

Cat. No. 10 236 250 001 y Version 06 ␮ c 50 g 1 A260 unit Content version: September 2018 50 ␮g (200 ␮l) for 50 gel lanes Store at Ϫ15 to Ϫ25°C

Product overview

Formulation Solution in 10 mM Tris-HCl, 1 mM EDTA, pH 8

Concentration 250 ␮g/ml

Size distribution The bacteriophage lambda (λ) DNA is fragmented in a restriction digestion with Hind III endonuclease. The digestion reaction results in 8 double-stranded DNA fragments: 23130, 9416, 6557, 4361, 2322, 2027, 564, and 125 as determined by computer analysis of the λDNA sequence.

Application DNA Molecular Weight Marker II provides accurate siz- ing of fragments over a broad range of sizes. The fragments have 5-protruding ends and can be labeled with radioactive nucleotides (e.g., [32P]-dTTP or [32P]-dGTP) by standard filling-in reactions. End-label- ing reactions can be performed with a radioactive or nonradioactive dideoxynucleotide (e.g., DIG-11-ddUTP*) and Terminal Transferase*.

Properties Upon electrophoretic separation, 7 bands are visible (see fig.) when resolved as described below. The small- est fragment resulting from the Hind III digestion (125 base pair) cannot be detected on the gel due to its small size. All fragments are present in equimolar amounts, therefore this smallest band will only be visible on over- loaded gels when stained with ethidium bromide. Fig.: Separation of 1 µg DNA Molecular Weight Marker II on Typical analysis The DNA fragment mixture shows the typical pattern a1% agarose MP gel, stained with ethidium bromide. with 7 bands in agarose gel electrophoresis, as shown in the figure. Ordering Information ␮ Separation Apply 1 g DNA per lane on a 1% Agarose MP gel. conditions Changes to Editorial changes. previous version Handling The 23130 and 4361 bp fragments contain the cos-ends instructions of lambda. These bands are visible after heating the Regulatory For life science research only. Not for use in diagnostic marker at 65°C for 10 minutes, and quickly chilling on Disclaimer procedures. ice (see note below). Under standard conditions using ethidium bromide, the 125 bp fragments is visible only Disclaimer of For patent license limitations for individual products on over-loaded agarose gels. License please refer to: List of biochemical reagent products For higher resolution, we recommend to use 0.35 ␮g per gel well in a 0.6% (or lower) gel with 4 mm thickness. The fragment of 564 base pairs will then be easily visible. Note: General comment about separation conditions of digested λDNA molecular weight markers concerning the cos-ends of lambda: Fragments containing the 12 base cos-sites of lambda may anneal upon storage. This leads to a gel pattern where one band is of lower intensity than expected (or absent completely) and a larger fragment has an increased intensity. Denaturation of the cos-sites can be Contact and Support performed immediately before loading the gel, by heat- ing at 65°C for 10 minutes and quick-chilling on ice. To ask questions, solve problems, suggest enhancements and report new applications, please visit our Online Technical Support Site. Ϫ Ϫ Storage/stability The unopened vial is stable at 15 to 25°C until the To call, write, fax, or email us, visit sigma-aldrich.com, and select your home expiration date printed on the label country. Country-specific contact information will be displayed. Note: This product is shipped in dry ice and should be stored at Ϫ15 to Ϫ25°C until use. Once thawed we rec- ommend further storage at + 2 to + 8°C. Repeated freezing and thawing should be avoided.

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