Objectives

• Compare and contrast among the following in vitro assays for amplifying nucleic acids: polymerase chain reaction (PCR), branched DNA amplification, ligase chain reaction, transcription-mediated amplification, and QP replicase with regard to type of target nucleic acid, principle, major elements of the procedure, type of amplicon produced, major enzyme(s) employed, and applications.

• Describe examples of modifications that have been developed for PCR.

• Discuss how amplicons are detected for each of the amplification methods.

• Design forward and reverse primers for a PCR, given the target sequence.

• Differentiate between target amplification and signal amplification.

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