Specific Dna Amplification
Specific Dna Amplification - Related Videos
09:31
Amplification, Next-generation Sequencing, and Genomic DNA Mapping of Retroviral Integration Sites
0 Views •
Cited by 56 •
2016
We describe a protocol for amplifying retroviral integration sites from the genomic DNA of infected cells, sequencing the amplified virus-host junctions, and then mapping these sequences to a reference genome. We also describe techniques to quantify the distribution of integration sites relative to various genomic annotations using BEDTools.
09:36
Visualization of Mitochondrial DNA Replication in Individual Cells by EdU Signal Amplification
0 Views •
Cited by 11 •
2010
We developed a sensitive technique to label newly synthesized mitochondrial DNA (mtDNA) in individual cells in order to study mtDNA biogenesis. The technique combines the incorporation of EdU together with a tyramide signal amplification (TSA) protocol to visualize mtDNA replication within subcellular compartments of neurons.
09:44
Rapid Amplification of cDNA Ends
0 Views •
2023
Source: Pablo Sanchez Bosch2, Sean Corcoran2 and Katja Brückner1,2,3 1Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research 2Department of Cell and Tissue Biology, 3Cardiovascular Research Institute, University of California San Francisco, San Francisco, CA, USA Rapid Amplification of cDNA Ends (RACE) is a technique that allows amplification of full-length cDNA from mRNA by extending to the 3’ or 5’ end, even without prior knowledge of the sequence (Frohman et al.,...
11:58
Linear Amplification Mediated PCR – Localization of Genetic Elements and Characterization of Unknown Flanking DNA
0 Views •
Cited by 14 •
2014
Linear-amplification mediated (LAM)-PCR is a method developed to identify the exact positions of integrating viral vectors in the genome. The technique has evolved to be the superior method to study clonal dynamics in gene therapy patients, biosafety of novel vector technologies, T-cell diversity, cancer stem cell models, etc.
06:57
Loop-mediated Isothermal Amplification (LAMP) Assays for the Species-specific Detection of Eimeria that Infect Chickens
0 Views •
Cited by 10 •
2015
Diagnosis of Eimeria infection in chickens remains demanding. Parasite morphology- and host pathology-led approaches are commonly inconclusive, while molecular approaches based on PCR have proven demanding in cost and expertise. The aim of this protocol is to establish loop-mediated isothermal amplification (LAMP) as a straightforward molecular diagnostic for eimerian infection.
FAQs
Primer placement determines the boundaries of the product. The two primers bind to sequences flanking the chosen region, so DNA synthesis proceeds across the intervening segment rather than indiscriminately copying the entire sample. This arrangement gives specific DNA amplification its selectivity and allows researchers to examine a defined genetic region within a complex mixture.
Each cycle separates the DNA strands, allows the primers to bind their complementary sequences, and lets the thermostable polymerase extend from those primers. Newly made strands then serve as templates in later cycles. Because the target is copied repeatedly, successive cycles produce exponential enrichment, making small amounts of the selected sequence easier to analyze.
The thermostable DNA polymerase carries out extension after the DNA strands have been separated by denaturation. Its stability allows it to remain functional through repeated cycles, supporting continuous copying of the primer-defined region. Without a polymerase that tolerates these cycling conditions, the sequence-specific amplification process could not efficiently generate the enriched DNA product.
Target selection focuses analysis on one defined sequence instead of treating all DNA in the sample as equally informative. Primers provide that selection by identifying the flanking sites, while repeated copying enriches the intervening region. This focused approach is especially useful when the desired sequence represents only a small fraction of a complex biological sample.
A basic workflow follows the cycle sequence of denaturation, primer annealing, and extension. Denaturation separates the DNA strands, annealing positions the primers at the flanking sequences, and extension allows the thermostable polymerase to copy the target region. Repeating these stages progressively enriches the selected product for downstream genetic analysis.
The method is useful when researchers need sensitive examination of a particular DNA sequence in a biological sample. In pathogen identification, target selection can support detection of pathogen-associated genetic material. In genotyping and mutation analysis, the enriched region provides material for characterizing sequence-specific genetic differences rather than analyzing the entire sample indiscriminately.
Specific DNA amplification can prepare a defined genetic region for subsequent use in cloning or sequencing. By enriching the sequence between the primers, the method produces more material for characterization or incorporation into a cloning workflow. This makes it valuable as a sample-preparation step when the original biological material contains insufficient amounts of the region of interest.