Sanger sequencing method

The Sanger sequencing method is outlined here. The principle depends on the ability of an enzyme, DNA polymerase, to make a very accurate complementary... 

Figure 28.8 The sequence of a restriction fragment determined by the Sanger dideoxy method can be read simply by noting the colors of the dye attached to each of the various terminal nucleotides.

Sequencing of DNA is carried out by the Sanger dideoxy method, and small DNA segments can be synthesized in the laboratory by automated instruments. Small amounts of DNA can be amplified by factors of 106 using the polymerase chain reaction (PCR). 

Sanger dideoxy method (Section 2.6) The most commonly used method of DNA sequencing. 

The amino acid sequences of thousands of different proteins from many species have been determined using principles first developed by Sanger. These methods are still in use, although with many variations and improvements in detail. Chemical protein sequencing now... 

FIGURE 3-25 Steps in sequencing a polypeptide, (a) Identification of the amino-terminal residue can be the first step in sequencing a polypeptide. Sanger s method for identifying the amino-terminal residue is shown here, (b) The Edman degradation procedure reveals... 

Correct answer = B. The sequence of the new strand of DNA synthesized using the Sanger dideoxy method may be determined by reading the bands from the bottom to the top of the gel as 5TACCAG3 This is complementary to the original strand, which is. therefore, 5 CTGGTA3 ... 

Common method of DNA sequencing Cloned, purified fragments of DNA can be sequenced using the Sanger dideoxy method. 

The Sanger dideoxynucleoside method of sequencing DNA. (a) A suitable template is chosen, and the primer is chosen so that DNA synthesis begins at the point of interest. The primer is radioactively labeled. In addition to the template-primer complex the reaction mixture contains all four radioactive deoxyribonucleoside triphosphates and small amounts of a single dideoxynucleoside triphosphate. The dideoxy compound serves as a chain terminator. 

Sanger and Barrell developed rapid DNA-sequencing methods. 

This technique uses a combination of SSCP and Sanger sequencing to create a larger set of bands for analysis in a native acrylamide gel (7). In this method... 

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