Nucleic acid sequencing Maxam-Gilbert

The enormous advances in nucleic acid sequencing (Maxam and Gilbert, 1977 Sanger et al., 1977, and earlier papers) and their relative simplicity and ease as compared with amino acid sequencing not only should provide confirmation of amino acid sequences of V-regions but may ultimately provide additional sequences of known antibodies, especially when carried out on antibodies of known specificity from hybridomas. 

Fig. 3. Nucleic acid sequencing. Scheme showing the chemical cleavage method of Maxam and Gilbert applied to the determination of the nucleotide sequence in a hypothetical length of DNA. a = original DNA, i.e. the uncleaved but P-labeled DNA being sequenced, b > location of each nucleotide in the DNA being sequenced, numbering from the 5 -end.

In this chapter, the Maxam-Gilbert method and the Sanger method for ab initio sequencing of long nucleic acid chains are discussed. Two alternative methods for sequencing of shorter DNA strands are described in chapter 5 with DNA arrays, sequences of up to a few hundred base pairs can be determined (section 5.3), while pyrosequencing is capable of identifying sequences of up to 50 bp (section 5.4). 

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