Home  Contacts  Order  Catalog  Support  Search  Alphabetical Index  Numerical Index  Restriction Enzymes  Modifying Enzymes  PCR, qPCR, RT-PCR & dNTPs  Molecular Cloning  Nucleic Acid Purification  Molecular Labeling & Detection  In vitro Transcription  Electrophoresis Products  Nucleotides  Transfection Reagents  Reagents C A T A L O G

6X DNA Loading Dye & SDS Solution

#R1151	5x1 ml
Related Documents (in pdf, ~30-50 KB): Certificate of Analysis: #R1151 MSDS (English) MSDS (English-USA) MSDS (German)

 

Features

• Visual two-color tracking of DNA migration during electrophoresis.

• No DNA masking during gel exposure to UV light.

Figure 1. Electrophoresis of tracking dyes in 6X DNA Loading Dye & SDS Solution.

Figure 2. The effect of SDS for electrophoresis of DNA samples containing high amounts of DNA binding proteins. M - GeneRuler™ DNA Ladder Mix 1 - 0.5 µg lambda DNA with 6X DNA Loading Dye 2 - 0.5 µg lambda DNA with 6X DNA Loading Dye & SDS Solution 3 - 0.5 µg lambda DNA/TsoI with 6X DNA Loading Dye 4 - 0.5 µg lambda DNA/TsoI with 6X DNA Loading Dye & SDS Solution 5 - 0.4 µg of the 2 fragment mixture prior the addition of T4 DNA Ligase 6 - 0.4 µg of the 2 fragment mixture after the ligation with T4 DNA Ligase, with 6X DNA Loading Dye 7 - 0.4 µg of the 2 fragment mixture after the ligation with T4 DNA Ligase, with 6X DNA Loading Dye & SDS Solution

Description
The 6X DNA Loading Dye & SDS Solution is recommended for preparation of DNA samples with high amounts of DNA binding proteins prior to loading on agarose and polyacrylamide gels. It is supplemented with 1% SDS to eliminate DNA-protein interactions. Proteins have to be denatured to release DNA from DNA-protein complexes. Otherwise, the DNA binding proteins may alter electrophoretic DNA migration, cause DNA samples to stick to gel wells or band shifts. SDS also prevents the annealing of DNA molecules via long cohesive ends and helps to avoid additional bands in electrophoresis. The optimized 6X DNA Loading Dye & SDS Solution contains two electrophoresis tracking dyes: bromophenol blue and xylene cyanol FF (see Fig.1). It also contains 100 mM EDTA, that binds divalent metal ions, inhibits metal dependent enzymatic reactions and protects DNA from metal dependent nucleases, effectively inactivating enzymes and protecting DNA from nucleases.

Applications

• Recommended for electrophoretic analysis of DNA samples with high amounts of proteins.

• Prevents appearance of additional bands due to annealing of DNA molecules with cohesive ends.
• Recommended for kinetic experiments - 100 mM EDTA ensures immediate stopping of metal dependent enzymatic reactions.
• Recommended for DNA agarose gel analysis after DNA restriction digestions, ligation or dephosphorylation reactions.

Composition of 6X Solution

• 0.03% bromophenol blue

• 0.03% xylene cyanol FF
• 60% glycerol
• 1% SDS
• 100 mM EDTA (pH 7.6, adjusted with Tris)

Note

• In 1% agarose gels bromophenol blue co-migrates with ~300 bp DNA, while xylene cyanol FF co-migrates with ~4000 bp DNA.

• For the migration rates of dyes in agarose and polyacrylamide gels see tables.
• The prepared sample can be stored at -20°C. It can be subjected and is viable for several freeze-thaw cycles.

Quality Control
Tested for DNA sample preparation prior to agarose gel electrophoresis.

Storage
Store at room temperature or at +4°C up to 12 months.
For longer periods, store at -20°C.

Usage Recommendations

Add 1 volume of 6X DNA Loading Dye & SDS Solution to 5 volumes of DNA sample. Mix well. Heat at 65°C for 10 minutes. Chill on ice, spin down and load.

 

Related Products

TopVision™ LE GQ Agarose TopVision™ LM GQ Agarose 10X TBE Buffer 50X TAE Buffer Agarase DNA Markers/Ladders DNA Gel Extraction Kit

Updated vasario 01, 2008 09:30