Multiplex ligation-dependent probe amplification is a PCR-based method whereby two probes that hybridise adjacent to each other on the target DNA are joined by a ligase enzyme and amplified using a single primer by virtue of generic tails synthesized onto each set of probes. Each probe set is designed to produce a product of a predetermined length which allows differentiation on the basis of size. A fluorescent dye is attached to one of the PCR primers allowing for detection by capillary electrophoresis, if desired.
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MBiTMLPA-based Binary Typing (MBiT) is a system of subtyping bacteria based on the presence or absence of each of a range of genetic targets hypothesised to be involved in pathogenicity or survival. It is a single-reaction version of the PCR-based Binary Typing (P-BIT) system designed by ESR for the subtyping of foodborne pathogens. It is inexpensive, rapid, highly portable and requires only basic molecular biology equipment.
The presence or absence of each target is scored as 1 or 0 to make a binary code. This code can then be shortened to a smaller string of numbers by dividing the targets into groups of three, multiplying the result for the first target in the group by 1, the second by 2 and the third by 4. The results for each group are added together to give a group score of between 0 and 7. The group scores are then concatenated.
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