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Maxam–Gilbert sequencing is a method of DNA sequencing developed by Allan Maxam and Walter Gilbert in 1976–1977. This method is based on nucleobase-specific partial chemical modification of DNA and subsequent cleavage of the DNA backbone at sites adjacent to the modified nucleotides. [1] An example Maxam–Gilbert sequencing reaction.
Walter Gilbert, a biochemist, and Allan Maxam, a molecular geneticist, at Harvard also developed sequencing methods, including one for "DNA sequencing by chemical degradation". [39] [40] In 1973, Gilbert and Maxam reported the sequence of 24 basepairs using a method known as wandering-spot analysis. [41]
Allan Maxam and Walter Gilbert’s 1977 paper “A new method for sequencing DNA” was honored by a Citation for Chemical Breakthrough Award from the Division of History of Chemistry of the American Chemical Society for 2017. It was presented to the Department of Molecular & Cellular Biology, Harvard University. [4] [1]
The method uses an enzyme, deoxyribonuclease (DNase, for short), to cut the radioactively end-labeled DNA, followed by gel electrophoresis to detect the resulting cleavage pattern. For example, the DNA fragment of interest may be PCR amplified using a 32 P 5' labeled primer, with the result being many DNA molecules with a radioactive label on ...
In January 1978, David J. Galas and Albert Schmitz developed the DNA footprinting technique to study the binding specificity of the lac repressor protein. It was originally a modification of the Maxam-Gilbert chemical sequencing technique. [1] The method was submitted and published without revision in Nucleic Acids Research.
Sanger's approach was described in 2001 as one of the two fundamental methods for sequencing DNA fragments [1] (the other being the Maxam–Gilbert method [5]) but the Sanger method is both the "most widely used and the method used by most automated DNA sequencers."
The first DNA sequencing methods were developed by Gilbert (1973) [8] and Sanger (1975). [9] Gilbert introduced a sequencing method based on chemical modification of DNA followed by cleavage at specific bases whereas Sanger's technique is based on dideoxynucleotide chain termination. The Sanger method became popular due to its increased ...
[58] [59] High-throughput sequencing is intended to lower the cost of DNA sequencing beyond what is possible with standard dye-terminator methods. In ultra-high-throughput sequencing, as many as 500,000 sequencing-by-synthesis operations may be run in parallel. [60] [61] Illumina Genome Analyzer II System.