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Electrophoresis Articles

RNA Electrophoresis

By National Diagnostics | September 2, 2011 | Comments Off on RNA Electrophoresis

Electrophoretic analysis of RNA presents unique challenges. RNA is isolated in single-stranded form, without complementary sequences. It must be fully denatured in order to obtain fractionation based on size. However, RNA molecules form complex and…

PFGE and FIGE

By National Diagnostics | August 31, 2011 | Comments Off on PFGE and FIGE

Standard agarose gels can resolve DNA fragments up to 75 kb. Often, analysis of genomic DNA requires resolution of megabase (mb) fragments. Fragments greater in size than one megabase all run at the same rate…

In Gel Enzyme Reactions

By National Diagnostics | August 31, 2011 | Comments Off on In Gel Enzyme Reactions

In many cases, the processing of DNA by enzymes is not impeded by agarose. Such reactions can be run directly in bands excised from low melting point agarose gels. The excised band is melted, mixed…

DNA/RNA Purification from Agarose Gels – Electroelution

By National Diagnostics | August 31, 2011 | Comments Off on DNA/RNA Purification from Agarose Gels – Electroelution

The most popular alternative to glass powder elution for the complete purification of DNA from agarose is electroelution. Because agarose gels are run in a horizontal apparatus, the gel can be manipulated during a pause…

Restriction Digest Mapping

By National Diagnostics | August 29, 2011 | Comments Off on Restriction Digest Mapping

Restriction endonucleases are enzymes that cleave double-stranded DNA at specific sites, generally 4, 6 or 8 base palindromic sequences. Because, in action, the enzymes are sequence-specific, each piece of DNA has a recognizable pattern (or…

Preparation of Denaturing Agarose Gels

By National Diagnostics | August 29, 2011 | Comments Off on Preparation of Denaturing Agarose Gels

A variety of denaturants can be used with agarose. Alkaline gels are most often employed with single-stranded Preparing Alkaline Agarose Gels At high temperatures, alkaline conditions will hydrolyze the Agarose polysaccharide chains. To prepare an…

Preparation of Agarose Gels

By National Diagnostics | August 29, 2011 | Comments Off on Preparation of Agarose Gels

Use the tables below corresponding to the AquaPor agarose of choice to determine the percentage gel to cast. (LE = general-purpose; LM = low melting; ES = extra strength; HR = high resolution). AquaPor LE…

Agarose Gel Electrophoresis of DNA and RNA – Uses and Variations

By National Diagnostics | August 26, 2011 | Comments Off on Agarose Gel Electrophoresis of DNA and RNA – Uses and Variations

Agarose electrophoresis is used for a variety of purposes. Specialty grades of agarose have been developed to fulfill specific requirements. The most commonly used variant is low-melting agarose, which has been modified to lower its…

Agarose Gel Electrophoresis of DNA and RNA – An Introduction

By National Diagnostics | August 26, 2011 | Comments Off on Agarose Gel Electrophoresis of DNA and RNA – An Introduction

DNA and RNA strands are extremely large macromolecules. A 1 kilobase piece of single-stranded DNA or RNA has a molecular weight of 330,000 daltons, larger than the vast majority of proteins. Often in the molecular…

SSCP Analysis

By National Diagnostics | August 26, 2011 | Comments Off on SSCP Analysis

Single-stranded DNA can adopt multiple conformations under non-denaturing conditions. In the absence of a complementary strand, DNA will anneal short internal complementary sequences, forming a complex “knot.” A DNA molecule follows a complex path of…