Dideoxy sequencing technique

Rapid sequencing techniques have been developed to further the analysis of DNA molecules. DNA can be sequenced by controlled interruption of replication (Sanger dideoxy method). The fragments produced are separated by gel... 

The dideoxy fingerprinting (ddF) is described by Sarkar et al. (92) to be a hybrid between dideoxy sequencing and SSCP. The method is rapid, large and small regions can be amplified and screened. In the initial publication, Sarkar et al. detected 84 out of 84 known mutations. The frequency of false positivity has been described to be low—in the order of 5% (92,97). The ddF technique has been explored on 73 primary breast cancers (97). Sarkar et al. claimed that this technique detected 100% of the gene mutations, but compared with SSCP, ddF technique requires 50% more effort (97). 

Section 28 14 The nucleotide sequence of DNA can be determined by a technique m which a short section of single stranded DNA is allowed to produce its complement m the presence of dideoxy analogs of ATP TTP GTP and CTP DNA formation terminates when a dideoxy analog is incorporated into the growing polynucleotide chain A mixture of polynucleotides dif fermg from one another by an incremental nucleoside is produced and analyzed by electrophoresis From the observed sequence of the comple mentary chain the sequence of the original DNA is deduced... 

After removal of the plasmid from the phage, the plasmid can be used in reaction mixtures used for DNA sequencing. The dideoxy technique for sequencing DNA is shown below. 

With use of the dideoxy technique, and with viewing the autoradiographic gel (the "aulorad", would you expect to be able to read and determine the DNA sequence that occurs in the primer ... 

Small DNA segments can be synthesized in the laboratory, and commercial instruments are available for automating the work. Sequencing of DNA can be carried out either by the Maxam-Gilbert method, which uses chemical techniques, or by the Sanger dideoxy method, which uses enzymatic techniques. Small amounts of DNA can be amplified by factors of 10 using the polymerase chain reaction (PCR). 

Southern and Northern blotting techniques (p. 137) Sanger dideoxy method of DNA sequencing (p. 138) Solid-phase synthesis of nudeic acids (p. 139)... 

Which of the following sets of reagents are required for dideoxy chain DNA synthesis in the Sanger technique for DNA sequencing . ... 

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