Microsatellite Amplification
Microsatellite Amplification - Related Videos
09:44
Rapid Amplification of cDNA Ends
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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.,...
10:12
Protein Misfolding Cyclic Amplification of Prions
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Cited by 7 •
2012
Protein misfolding cyclic amplification (PMCA) is an in vitro assay for the study of prion conversion and strain and species barriers. It can also be used as a prion detection assay.
02:41
Amplification of a Suicide Plasmid in a Bacterial Host
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2025
Source: Tomás, J. M., et al. Generation of Null Mutants to Elucidate the Role of Bacterial Glycosyltransferases in Bacterial Motility. J. Vis. Exp. (2022).This video demonstrates the initiator-dependent amplification of a suicide plasmid in E. coli. The plasmid, which carries an antibiotic resistance gene, replicates only in the presence of an initiator protein encoded by the host. After electroporation and incubation in recovery medium, the mixture is plated on antibiotic-containing agar.
02:48
Loop-Mediated Isothermal Amplification for Bacterial Gene Detection
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2025
Source: Domesle, K. J., et al. Loop-Mediated Isothermal Amplification for Screening Salmonella in Animal Food and Confirming Salmonella from Culture Isolation. J. Vis. Exp. (2020).This video demonstrates the loop-mediated isothermal amplification (LAMP) technique for detecting a target bacterial gene by amplifying DNA at a constant temperature. The reaction mixture is prepared by adding an isothermal master mix, specific primers for the target gene, and samples containing DNA templates. The...
07:35
Demonstrating a Multi-drug Resistant Mycobacterium tuberculosis Amplification Microarray
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Cited by 4 •
2014
An amplification microarray combines asymmetric PCR amplification and microarray hybridization into a single chamber, which significantly streamlines microarray workflow for the end user. Simplifying microarray workflow is a necessary first step for creating microarray-based diagnostics that can be routinely used in lower-resource environments.
FAQs
Primers are designed to bind on the DNA sequences flanking the tandem repeat, so amplification includes the variable region between them. Their placement allows the resulting product length to reflect differences in repeat size among samples. Primer specificity is therefore central to obtaining a clear target rather than a mixture of unrelated DNA fragments.
Each cycle separates the DNA strands during denaturation, allows primers to bind during annealing, and uses DNA synthesis to copy the region between the primers. Repeating these stages produces millions of target copies from the original sample. The cycle sequence makes small length differences in the amplified fragments measurable after separation or detection.
Nonspecific products arise when amplification generates DNA fragments other than the intended microsatellite target. They can complicate fragment separation and make allele sizing less reliable, especially when products overlap or obscure the expected signal. Careful primer selection and control of amplification conditions help preserve the specificity needed for comparing fragment lengths among samples.
After amplification, fragments can be separated by electrophoresis, which resolves DNA products according to their lengths, or analyzed through fluorescent detection. The measured fragment sizes indicate differences in the amplified repeat region. Reliable interpretation depends on consistent allele sizing, because the biological comparisons rely on distinguishing small length differences among individuals or samples.
The workflow begins with primers that flank the selected repeat region, followed by PCR cycles that copy the target. The amplified products are then examined using electrophoresis or fluorescent detection, and their fragment sizes are compared across samples. This sequence converts variation in repeat length into data suitable for genetic profiling or other biological analyses.
Its high variability makes the method useful for genetic profiling, population studies, linkage mapping, biodiversity assessments, and forensic identification. Researchers can compare fragment lengths among individuals or samples to examine genetic differences and relationships. The appropriate application depends on whether the goal is tracking variation, studying inheritance, assessing biodiversity, or distinguishing biological samples.