Polymerase chain reaction probe sequences

LU c c, TANG K F J, KOU G H and CHEN s N (1993), Development of a Penaeus monodon-type baculovirus (MBV) DNA probe by polymerase chain reaction and sequence analysis, J Fish Dis, 16, 551-559. 

A variation on the theme of conventional assay uses both lanthanide-labeled and biotin-labeled single strands to form split probes for sequence of target strands (Figure 12).120 When both of these bind to DNA, the complex binds (via the biotin residue) to a surface functionalized with streptavidin, immobilizing the europium and allowing assay to be carried out. This approach is already very sensitive to DNA sequence, since both sequences must match to permit immobilization of the lanthanide, but can be made even more sensitive by using PCR (the polymerase chain reaction) to enhance the concentration of DNA strands. In this way, initial concentrations corresponding to as few as four million molecules can be detected. This compares very favorably with radioimmunoassay detection limits. 

Format 1 SBH can be used to uncover polymorphism and mutations in a particular gene. The sample amplicons of genomic DNA of a test individual and the amplicons for the control DNA for the gene of interest with a known reference sequence are both prepared by polymerase chain reaction (PCR). A subset of probes that is complementary... 

Such renaturation or annealing of complementary DNA strands is an important step in probing a Southern blot and in performing the polymerase chain reaction (reviewed in Chapter 7). In these techniques, a well-characterized probe DNA is added to a mixture of target DNA molecules. The mixed sample is denatured and then renatured, When probe DNA binds to target DNA sequences of sufScient complementarity, the process is called hybridization. 

Nucleic acid-based technologies Nucleic acid probe Polymerase chain reaction, DNA amplication 16S rRNA sequencing techniques automated riboprinting... 

As the genes themselves are identified and sequenced, so the need for RFLP markers declines since specific DNA probes (see Topic 12) for the most common types of gene defect can be employed. In addition, the use of the polymerase chain reaction (PCR see Topic 16) in screening for human genetic disease is becoming the method of choice rather than RFLP analysis since it is much faster to perform and requires far less clinical material for analysis. 

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