DNA Electrophoresis in Agarose Gels Protocols

Protocol for agarose gel electrophoresis of DNA. Includes: Making the gel; Loading the gel; Running the gel.
- [Read Agarose Gel Electroporesis of DNA Protocol]

Alkaline agarose gels are used chiefly to measure the size of first and second strands of cDNA (Construction of cDNA Libraries Stage 1: Synthesis of First-strand cDNA Catalyzed by Reverse Transcriptase) and to analyze the size of the DNA strand after digestion of DNA-RNA hybrids with nucleases such as S1. - [Read Alkaline Agarose Gel Electrophoresis Protocol]

The most convenient and commonly used method to visualize DNA in agarose gels is staining with the fluorescent dye ethidium bromide.  Ethidium bromide can be used to detect both singleand double-stranded nucleic acids (both DNA and RNA).  However, the affinity of the dye for single-stranded nucleic acid is relatively low and the fluorescent yield is comparatively poor. - [Read Detection of DNA in Agarose Gels Protocol]

Protocol for electrophoresis of single stranded DNA using agarose gel. - [Read Electrophoresis - Agarose Gels for Single Stranded DNA Protocol]

A messy but reliable technique that works well for DNAs ranging in size from 200 bp to >50 kb. - [Read Recovery of DNA from Agarose and Polyacrylamide Gels: Electroelution into Dialysis Bags Protocol]

Recovery of bands of DNA from agarose gels by electrophoresis onto a sliver of DEAE-cellulose membrane can be performed simultaneously on many samples and reliably gives high yields of fragments between 500 bp and 5 kb in length. - [Read Recovery of DNA from Agarose Gels: Electrophoresis onto DEAE-cellulose Membranes Protocol]

A fragment of gel containing a band of DNA is excised and digested with agarase, which hydrolyzes the polymer to disaccharide subunits.  The released DNA is then purified by phenol extraction and ethanol precipitation.  The method works well for DNAs ranging in size from <5 kb to >20 kb. - [Read Recovery of DNA from Low-melting-temperature Agarose Gels: Enzymatic Digestion with Agarase Protocol]

DNA fragments separated by electrophoresis through gels cast with low-melting-temperature agarose are recovered by melting the agarose and extracting the resulting solution with phenol:chloroform.  The protocol works best for DNA fragments ranging in size from 0.5 kb to 5 kb. - [Read Recovery of DNA from Low-melting-temperature Agarose Gels: Organic Extraction Protocol]