Mugpcr

Two tube

Universal genotyping platform

[17]

Abbreviations: ADPL=allele discrimination by primer length; PAMSA=PCR amplification of multiple specific alleles; CB = competitive blocker; CTPP=confronting two-pair primer; MUG=multiplex universal genotyping.

Abbreviations: ADPL=allele discrimination by primer length; PAMSA=PCR amplification of multiple specific alleles; CB = competitive blocker; CTPP=confronting two-pair primer; MUG=multiplex universal genotyping.

allele-specific reaction internal control reaction

Fig. 1 Principle of ARMS-PCR (see text for details). (From Vamvakopoulos, J.E., 2001. A potential role for interleukin-1 in chronic allograft rejection. PhD thesis. University of Cambridge, U.K.) (View this art in color at www.dekker.com.)

allele-specific reaction internal control reaction

Fig. 1 Principle of ARMS-PCR (see text for details). (From Vamvakopoulos, J.E., 2001. A potential role for interleukin-1 in chronic allograft rejection. PhD thesis. University of Cambridge, U.K.) (View this art in color at www.dekker.com.)

further action should involve modification of the allele-specific primers[20,22,25,27-29] or use of depository oligos.[30] ARMS-PCR relying on intercalating fluorescent dyes for amplicon detection may require additional troubleshooting aimed at minimizing nonspecific fluorescence from primers and primer dimers. This issue becomes crucial for applications demanding optimal sensitivity, such as genotyping of DNA pools and monitoring minimal residual disease or the emergence of drug-resistant infectious strains in clinical samples. Primer design measures for minimizing primer dimer formation have been described;[31] alternatively, optimal signal-to-noise ratio in these instances can be achieved by simply raising the reaction pH to 11.7 before amplicon detection.[32]

APPLICATIONS OF ARMS-PCR: GENOTYPING AND BEYOND

The versatility and success of ARMS-PCR as a methodological tool can best be judged by its uses. Here I summarize some of the most important, diagnostically oriented applications of this technique.

Clinical Genetics and Cancer Management

ARMS-PCR was originally conceived mainly for the purpose of detecting cancer-related mutations[1] and for studying the genetic basis of heritable disorders.[2-6] In this respect the field of clinical genetics was quick to explore the potential of this technique: rapid, PCR-based assays for several genetic diseases where the causative mutation was known were developed soon after the first description of ARMS-PCR.[33,34] Furthermore, the power of ARMS-PCR to detect mutations with unparalleled sensitivity has revolutionized two key aspects of cancer management, namely molecular diagnosis1-35-1 and the monitoring of minimal residual disease (MRD).[36,37]

Tissue Typing and Cell Therapeutics

HLA typing was one of the very first applications of ARMS-PCR and, in retrospect, the impact of this technique on histocompatibility testing has been momentous. Fugger et al.[38] pioneered the application of ARMS-PCR in this field, while Olerup, Zetterquist, and co-workers are credited with much of the work leading to the routine clinical application of this technique.[39,40] Today, ARMS-PCR is the bedrock of HLA typing. With new HLA specificities being integrated as they are discov-ered,[41] this situation looks set to persist long into the future.

Also of clinical relevance is the application of ARMS-PCR to the functional and quantitative study of cellular chimerism following cell or solid organ transplantation. Accurate quantitative measurement of cellular chimerism by ARMS-PCR is now feasible[42,43] and should greatly facilitate further advances in this field.

Studies of Interindividual Genetic Variation

The fact that ARMS-PCR is well suited to studying single nucleotide polymorphisms (SNPs), which account for over 80% of interindividual genetic variation, makes it an ideal tool for related functional and epidemiological studies. Besides conventional, biallelic SNPs, ARMS-PCR is easily adaptable to the study of triallelic SNPs,[44] small insertions, and deletions.[45] Where marker infor-mativeness becomes crucial (e.g., in mapping disease traits), this technique can also accommodate length polymorphisms alongside SNPs;[17] in that same context, it is also unique in allowing direct haplotyping of genetic markers.[17,46,47] Last but not least, the high sensitivity of this method makes it ideal for genotyping of pooled DNA samples.[48,49]

Getting Started With Dumbbells

Getting Started With Dumbbells

The use of dumbbells gives you a much more comprehensive strengthening effect because the workout engages your stabilizer muscles, in addition to the muscle you may be pin-pointing. Without all of the belts and artificial stabilizers of a machine, you also engage your core muscles, which are your body's natural stabilizers.

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