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. 2000 Jul;53(7):502–506. doi: 10.1136/jcp.53.7.502

Molecular techniques for clinical diagnostic virology

S Read 1, D Burnett 1, C Fink 1
PMCID: PMC1731222  PMID: 10961172

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Figure 1 Measurement of hepatitis B virus (HBV) genome in serum using real time monitoring of polymerase chain reaction (PCR) amplification kinetics of a set of standards on the LightCycler instrument. The PCR product is detected by a double stranded DNA specific fluorescent dye. Each line represents a PCR amplification. This experiment demonstrates the amplification of HBV DNA from a 1 log10 dilution series of the HBV DNA positive serum sample standard with an initial concentration of 107 genomes/ml. The concentration of HBV genome is proportionate to the PCR cycle at which the fluorescent signal increases above the background level (represented by the horizontal line).

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Figure 2 Additional information about the reaction detailed in fig 1 can be obtained from the melting curve analysis. Analysis of the melting characteristics of the polymerase chain reaction (PCR) products demonstrates two distinct products: primer dimer, melting at 74°C, and amplified hepatitis B virus (HBV) product melting at 84°C. Primer dimer formation occurs only at lowest input copy numbers.

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Figure 3 A "laboratory on a chip". A DNA chip containing immobilised arrays of oligonucleotides for the study of gene expression. Specialised equipment is required for interpreting the patterns generated by hybridisation of DNA to the array but the technique is suited to automated, high throughput applications.

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