Multiple-displacement amplification

Thompson, M. D., Bowen, R. A., Wong, B. Y., et al. (2005) Whole genome amplification of buccal cell DNA genotyping concordance before and after multiple displacement amplification. Clin. Chem. Lab. Med. 43, 157-162. 

Fig. 3. The use of microdissected material increased the sensitivity of the SNP array to LOH in tumor tissues. Regions with LOH are highlighted in black color. B, DNA extracted from bulk tumor tissue without microdissection M, DNA was extracted from microdissected tumor cells and amplified by the multiple displacement amplification method (52).

Instead of specific amplification of one target to improve sensitivity, methods that amplify all genomic DNA or mRNAs are useful when the target is in short supply. For example, multiple-displacement amplification uses exonuclease-resistant random hexamers and a highly pro-cessive polymerase to amplify DNA nonspecificaily. Initial DNA denaturation is not necessary and the reaction proceeds isothermally. Similarly, messenger RNA can be generi-caUy amplified with a poly(T) primer modified with an RNA polymerase promoter. After reverse transcription, second-strand DNA synthesis, and transcription, antisense RNA is produced. Both whole genome and antisense RNA amplification are also useful as nucleic acid purification methods before amplification or detection. 

Bray Ward P, et al. Comprehensive human genome amplification using multiple displacement amplification. Proc Natl Acad Sci U S A 2002 99 5261-6. 

Whole genome amplification of buccal cell DNA genotyping concordance before and after multiple displacement amplification. Clin Chem Lab Med 43 157-162... 

More recently, whole genome amplification (WGA), multiple displacement amplification (MDA) and other advanced copying techniques have been evolved [72]. 

---