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DNA Markers for Genomic DNA Analysis
For genomic DNA analysis by Southern blotting technique

Features

• Recommended for genomic DNA analysis in Southern blotting (see Protocol for Southern Blotting of Genomic DNA).
• No plasmid DNA sequences.
• Produce sharp bands.
• Broad range.
• The labeled marker probe can be used in a mixture with a test probe for genomic DNA hybridization.
• The markers do not hybridize with genomic DNA’s from Homo sapiens, Haemophilus influenzae, Bacillus subtilis, Physalis pruinosa, Saccharomyces cerevisiae.
• Bright bands of 3194 bp, 3976 bp and 5220 bp serve as references.
• Recommended for labeling with Klenow Fragment, exo- and by random-primed labeling with Fermentas DecaLabel™ and Biotin DecaLabel™ DNA Labeling Kits (see protocol).
• 50 ng of radioactively labeled marker is sufficient for 3-5 hybridization reactions.

Description & Other Information

Range Selection Guide

Product List:

• DNA Marker I for Genomic DNA Analysis (702-14321 bp)
• DNA Marker II for Genomic DNA Analysis (702-29946 bp)

Related Products

DNA Marker I for Genomic DNA Analysis

Designed for genomic DNA analysis by Southern blotting technique.
DNA Marker I for Genomic DNA Analysis is supplied with 10X DNA Loading Dye.

Catalog #	Certificate of Analysis (in pdf)	Product resources (in pdf)	Concentration, µg/µl	Amount, µg	Applications 50 ng/lane	Range,  bp	Fragments	Agarose,  %
#SM0341		MSDS (English) MSDS (English-USA) MSDS (German)	0.2	6	120	702-14321	28	0.7

50ng of the marker was run on a 20 cm length of 0.7% agarose gel in 1X TAE buffer at 3 V/cm 18 hours  (until bromophenol blue dye reached the bottom of the gel).  Blotting was carried out on SensiBlot™ Plus Nylon Membrane, and a [alfa32P] labeled marker was used as a probe for hybridization. Exposure time - 3 days. Range 28 fragments: from 702 to 14321 bp. Note The cohesive ends (the 12 nt cos site of lambda DNA) of some fragments may anneal and form additional bands.  These fragments can be separated by heating at 65°C for 5 min and then cooling on ice for 3 min.

DNA Marker II for Genomic DNA Analysis

Designed for genomic DNA analysis by Southern blotting technique.
DNA Marker II for Genomic DNA Analysis is supplied with 10X DNA Loading Dye.

Catalog #	Certificate of Analysis (in pdf)	Product resources (in pdf)	Concentration, µg/µl	Amount, µg	Applications 50 ng/lane	Range,  bp	Fragments	Agarose,  %
#SM0351		MSDS (English) MSDS (English-USA) MSDS (German)	0.2	6	120	702-29946	33	0.7

50ng of the marker was run on a 20 cm length of 0.7% agarose gel in 1X TAE buffer at 3 V/cm 18 hours  (until bromophenol blue dye reached the bottom of the gel).  Blotting was carried out on SensiBlot™ Plus Nylon Membrane, and a [alfa32P] labeled marker was used as a probe for hybridization. Exposure time - 3 days. Range 33 fragments: from 702 to 29946 bp. Note The cohesive ends (the 12 nt cos site of lambda DNA) of some fragments may anneal and form additional bands.  These fragments can be separated by heating at 65°C for 5 min and then cooling on ice for 3 min.

Description
Genomic DNA Markers I and II are specially designed for genomic DNA analysis by Southern blotting technique (see Protocol for Southern Blotting of Genomic DNA).
Both markers are prepared from lambda DNA and PhiX174 DNA; they do not possess any plasmid sequences. DNAs are separately digested to completion with the appropriate Fermentas PureExtreme® restriction endonuclease(s), purified, dissolved in a storage buffer and then mixed. The markers contain a mixture of blunt, and sticky ended DNA fragments. The markers are supplied with 10X DNA Loading Dye.

Concentration
0.2 µg/µl

Storage Buffer
10 mM Tris-HCl (pH 7.6) and 1 mM EDTA.

10X DNA Loading Dye
0.5% bromophenol blue, 50% glycerol and 100 mM EDTA

Recommendations for Loading

• Use the 10X DNA Loading Dye (supplied wth the marker) for your sample and marker DNA: mix 1 volume of the dye with 9 volumes of the DNA solution.
• Use ~6ng of the DNA Marker per 1 mm of an agarose gel lane width.
• Dilute 1 µl (0.2 µg) of DNA marker with 39 µl of nuclease-free water and mix.
• Prepare for loading:
  • 10 µl (50ng) DNA marker mix
  • 1.5 µl 10X DNA Loading Dye
  • water, nuclease-free up to 15 µl
• Vortex gently just prior to use.
• Heat DNA marker at 65°C for 5 min and chill on ice for 3 min before use.
• Apply the prepared amount (15 µl) of DNA marker on a 8 mm lane of agarose gel.

Quality Control
Tested in appropriate gel electrophoresis. The DNA concentration is determined spectrophotometrically. The absence of nucleases is confirmed by appropriate tests.

Storage
Store at -20°C.

Related Products

TopVision™ LE GQ Agarose TopVision™ LM GQ Agarose 50X TAE Buffer 10X TBE Buffer Agarase Klenow Fragment, exo- T4 Polynucleotide Kinase DNA Gel Extraction Kit Biotin DecaLabel™ DNA Labeling Kit DecaLabel™ DNA Labeling Kit Biotin-11-dUTP Biotin Chromogenic Detection Kit BCIP-T NBT SensiBlot™ Plus Nylon Membrane ATP 0.5 M EDTA, pH 8.0 Water, nuclease-free

Updated balandžio 24, 2008 18:26