By Rajyalakshmi Luthra, Rajesh Singh, Keyur Pravinchandra Patel
This up-to-date quantity explores a large choice of scientific functions of PCR similar to detecting DNA methylation, detection of viruses and protozoa in infectious ailments, estimation of gene replica quantity aberrations, primer extension coupled with mass spectroscopy, and excessive throughput NGS strategies. the applying of PCR has proven wonderful price within the learn of genomics and transcriptomics, not just for discovery but in addition for regimen medical functions, and it varieties the cornerstone of custom-made medication. Written for the hugely profitable Methods in Molecular Biology sequence, chapters contain short introductions to their respective issues, lists of the mandatory fabrics and reagents, step by step, effectively reproducible laboratory protocols, and pointers on troubleshooting and averting recognized pitfalls.
Thorough and authoritative, Clinical functions of PCR, 3rd Edition serves as a fantastic advisor for researchers aiming to appreciate the foundations at the back of each one software and for his or her implementation within the laboratory.
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Additional resources for Clinical Applications of PCR
Rabin KR, Man TK, Yu A et al (2008) Clinical utility of array comparative genomic hybridization for detection of chromosomal abnormalities in pediatric acute lymphoblastic leukemia. Pediatr Blood Cancer 51:171–177 11. Oostlander AE, Meijer GA, Ylstra B (2004) Microarray-based comparative genomic hybridization and its applications in human genetics. Clin Genet 66:488–495 12. Bejjani BA, Shaffer LG (2006) Application of array-based comparative genomic hybridization to clinical diagnostics. J Mol Diagn 8:528–533 13.
DdPCR droplet generation (DG) oil (Bio-Rad). 15. Bio-Rad QX100 droplet reader. 16. Droplet reader plate holders. 17. QuantaSoft software. 18. tec semi-skirted 96-well plate. 19. Heat sealer. 20. Heat sealing plate foil. 21. Thermal cycler. 3 Methods (Adapted from Jennings et al. ) 1. Incubate the RNA samples for 5 min at 75°. 2. Thaw the components of the One-Step RT-ddPCR kit from Bio-Rad Laboratories (Cat# 1863021; Hercules, CA) at room temperature. The contents of each of the reagents are adequately mixed by either pipetting or inverting followed by centrifugation.
It essentially combines the simplicity of traditional end-point PCR and the quantification features of the real-time quantitative PCR (qPCR) methodologies. Unlike qPCR, the quantitation is absolute and does not utilize calibration of standards, thus making the process faster, more precise, and reproducible . The basic principle involves extreme limiting dilution and partitioning of the sample into millions of separate units that ideally contain either no particle or a single particle. Each unit contains all the reagents necessary for a PCR reaction, and basically functions as a micro-PCR reactor.
Clinical Applications of PCR by Rajyalakshmi Luthra, Rajesh Singh, Keyur Pravinchandra Patel