Hybrid Primer
Hybrid Primer - Related Videos
11:14
Primer-Free Aptamer Selection Using A Random DNA Library
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Cited by 20 •
2010
SELEX protocols comprise multiple rounds of selection, each of which require regeneration of bound ligands, which in turn require fixed primer sequences flanking the random library regions. These fixed primer sequences can interfere with the selection process (false positives and negatives). Here we present a primer-free protocol.
08:00
Whole-Mount In Situ Hybridization
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2023
Whole-mount in situ hybridization (WMISH) is a common technique used for visualizing the location of expressed RNAs in embryos. In this process, synthetically produced RNA probes are first complementarily bound, or "hybridized," to the transcripts of target genes. Immunohistochemistry or fluorescence is then used to detect these RNA hybrids, revealing spatial and temporal patterns of gene expression. Unlike traditional in situ hybridization techniques, which require thin tissue sections whose...
15:28
Primer Extension Capture: Targeted Sequence Retrieval from Heavily Degraded DNA Sources
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Cited by 41 •
2009
We present a method of targeted ancient DNA sequence retrieval, which we used to reconstruct the complete mitochondrial genomes of five Neandertal individuals. Comparison of these sequences with present day humans suggests that Neandertals had a long term low effective population size.
03:24
Hybridization of Atomic Orbitals I
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2020
The mathematical expression known as the wave function, ψ, contains information about each orbital and the wavelike properties of electrons in an isolated atom. When atoms are bound together in a molecule, the wave functions combine to produce new mathematical descriptions that have different shapes. This process of combining the wave functions for atomic orbitals is called hybridization and is mathematically accomplished by the linear combination of atomic orbitals. The new orbitals that...
10:17
Laboratory Protocol for Genetic Gut Content Analyses of Aquatic Macroinvertebrates Using Group-specific rDNA Primers
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Cited by 7 •
2017
Most common gut content analyses of macroinvertebrates are visual. Requiring intense knowledge about morphological diversity of prey organisms, they miss soft bodied prey and, due to strong comminution of prey, are nearly impossible for some organisms, including amphipods. We provide detailed, novel genetic approaches for macroinvertebrate prey identification in the diet of amphipods.
FAQs
The complementary region provides the pairing step by annealing to the intended target strand. Once this alignment occurs, DNA polymerase extends the primer, copying the target-associated sequence into a new product. The distinct noncomplementary region can then become part of that product, linking target-specific amplification with the introduction of an additional designed sequence.
Although it does not initially pair with the target, the noncomplementary region is carried forward when DNA polymerase extends from the annealed portion of the primer. It can therefore become incorporated into newly synthesized DNA rather than remaining only a feature of the original oligonucleotide. This provides a mechanism for adding planned sequence information during amplification.
Target selectivity comes from the complementary region, because its sequence determines where the primer can anneal on the DNA template. The added region does not establish the initial target location; instead, it supplies the sequence intended for incorporation after binding occurs. Designing these regions together allows amplification to remain directed while the product receives a chosen modification.
The additional sequence gives the amplification product a function beyond simply reproducing the template region. Depending on its design, it can support the joining of genetic elements, introduce a site-directed change, or add an adapter or recognition sequence. Consequently, the same primer strategy can connect selective DNA amplification with a specific downstream engineering objective.
In PCR-based cloning, the complementary portion directs amplification of the selected DNA region, while the added portion contributes sequence needed for the intended construct. During polymerase extension, that extra sequence becomes incorporated into the amplified product. The resulting DNA can therefore combine a chosen target region with an engineered sequence for the cloning workflow.
For site-directed mutagenesis, the primer is designed so that amplification targets the relevant DNA region while the additional sequence introduces the planned change. Polymerase extension copies the primer-associated design into the product, allowing the amplified DNA to carry the modified sequence. This connects precise primer design with selective alteration of a chosen genetic element.
Beyond cloning and mutagenesis, hybrid primers can help create products that join separate genetic elements or contain added adapter and recognition sequences. These uses take advantage of the primer’s ability to combine template-directed binding with sequence addition. In biology research, that flexibility makes the approach relevant to broader genetic engineering and nucleic acid amplification workflows.