Dideoxy method of Sanger

A number of nucleotide analog compounds function by blocking further chain growth at the replication fork. The 2 3 -dideoxynucleosides can be converted to the triphosphates (ddNTPs). In the case of bacteria, these are incorporated onto the 3 -hydroxyl end of a growing DNA chain, and because the new end now lacks a 3 hydroxyl, no further additions can occur. They are used in conjunction with DNA polymerase I in the dideoxy method of Sanger for determining DNA sequences. 

The dideoxy method of Sanger for sequencing DNA involves the copying of a single strand of the DNA by a DNA polymerase (DNA polymerase I was originally used) to yield new strands (radioactively labeled for identification purposes), which are terminated at certain positions through the incorporation of a dideoxy nucleotide at the 3 end. A short primer, complementary to a sequence at one end of the single-strand template. 

Figure 0,7 depicts (he dideoxy method of DNA sequencing. This method was devised by Fred Sanger. The procedure is bipartite (i) utilization of piasmid DNA and DNApolymerase to create a few hundred different pieces of DNA, where each piece differs in Length (from the other pieces) by one nucleotide and (2) use of gel... 

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

Complete the following statements about the Sanger dideoxy method of DNA sequencing. 

The chain terminator method is also referred to as Dideoxy method or Sanger method. It is based on synthesising a complementary strand of DNA along the... 

In the chain termination or dideoxy method of DNA sequencing developed by Frederick Sanger, a primer-DNA template is divided into four separate reaction mixtures. To each is added the four dNTPs, one of which is labeled... 

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. 

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. 

Figure 5-21. DNA sequencing by the Sanger-dideoxy method. Single or double stranded DNA is used as a template for a primer-dependent DNA polymerase extension, which is carried out in four separate reaction mixes. In addition to DNA polymerase and the four dNTPs (including an a-radiolabelled nucleoside triphosphate), each reaction mix contains a small proportion of one dideoxy nucleotide derivative, ddATP,...

DNA Is Usually Sequenced by Controlled Termination of Replication (Sanger Dideoxy Method)... 

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