遗传学实验技术

A caged fluorescent nucleic acid probe, which contains a nucleotide modified with one photolabile nitrobenzyl unit and two hybridization-sensitive thiazole orange units, has been synthesized for area-specific fluorescence imaging of RNA in a cell. The probe emits very weak fluore ...

Significant efforts have been devoted to the development of techniques allowing the investigation of viral mRNA progression during the replication cycle. We herein describe the use of sequence-specific FIT–PNA (Forced Intercalation Peptide Nucleic Acids) probes which contain a ...

Molecular beacons are oligonucleotide (DNA or RNA) probes that have become increasingly important tools for RNA sensitive detection both in vitro and in living cells. From their inception, molecular beacons have been used to determine the expression levels of RNA transcripts, but they al ...

There is a lack of methods for high-throughput functional microRNA (miRNA) profiling. In this chapter, we describe a recombinant adeno-associated virus-based miRNA sensor array (miRNA Asensor array), which is able to profile functional miRNAs in cultured cells. The preparation of an mi ...

MicroRNAs play an important role in gene regulation and disease etiology and are blood-based biomarkers of diseases. Here, we describe a protein-facilitated affinity capillary electrophoresis (ProFACE) method for ultra-sensitive direct miRNA detection as low as 300,000 molec ...

Fluorescent in situ hybridization (FISH) is a powerful technology for studying the chromosome organization and aberrations as well as for searching the homology between chromosomal regions in mammals. Currently, FISH is used as a simple, rapid, and reliable technique for analyzing ch ...

Peptide nucleic acid (PNA) openers have the unique ability to invade double-stranded DNA with high efficiency and sequence specificity, making it possible to detect short (about 20 bp), single-copy bacterial DNA sequences. PNA openers bind to a target signature site on one strand of bacteri ...

Graphene oxide is chemically functionalized using planar structured first generation polyamidoamine dendrimer (G1PAMAM) to form graphene core GG1PAMAM. The monolayer of GG1PAMAM is anchored on the 3-mercapto propionic acid monolayer pre-immobilized onto a gold transduce ...

Optical label-free biosensors have demonstrated advantages over fluorescent-based detection methods by allowing accurate quantification while also being capable of measuring dynamic bimolecular interactions. A simple, high-throughput, solid-phase, and label-fr ...

Electrochemiluminescence (ECL) on bipolar electrode (BPE) is a sensitive, portable, and low-cost approach which has been employed to detect DNA and proteins. Here, we develop an ultrasensitive method for intracellular mRNA assay based on mRNA-mediated reporter DNA liberation and Ru ...

The detection of nucleic acid is of central importance for the diagnosis of genetic diseases, infectious agents, and biowarfare agents. Traditional strategies and technologies for nucleic acid detection are time-consuming and labor-intensive. Recently, isothermal strand-d ...

Many molecular biology experiments and clinical diagnostics rely on the detection or confirmation of specific nucleic acid sequences. Most DNA or RNA detection assays utilize radioactive or fluorescence labeling but although these tags are sensitive, safety issues (in the case of ra ...

We describe a method to detect DNA sequences visually through a color change reaction using DNAzymes. We successfully applied the assay for the detection of Salmonella and Mycobacterium DNA, as well as for genotyping single base differences from within human genomic DNA samples. Our appro ...

Although many approaches based on template replication were developed and applied in DNA detection, cross-contamination from amplicons is always a vexing problem. Thus, signal amplification is preferable for DNA detection due to its low risk of cross-contamination from amplicons. ...

Loop-mediated isothermal amplification (LAMP), which employs a Bst DNA polymerase with strand-displacement activity and four primers (two inner primers and two outer primers) recognizing six distinct regions on the target DNA, is a highly sensitive, specific, simple, and rapid nucl ...

Brucella spp. are facultative intracellular bacteria that cause zoonotic disease of brucellosis worldwide. Livestock that are most vulnerable to brucellosis include cattle, goats, and pigs. Brucella spp. cause serious health problems to humans and animals and economic losses to the ...

For patients infected with tuberculosis, detection of rpoB gene mutations is critical for diagnosing drug-resistant strains of the causative pathogen, Mycobacterium tuberculosis (MTB). Traditional approaches to drug resistance include culture, which is very slow. Recently ...

For effective control of pathogen-transmitting mosquitoes, precise surveillance data of mosquito distribution are essential. Recently, an increase of insecticide resistance due to the kdr mutation in Anopheles gambiae, a mosquito that transmits the malaria parasite, has been r ...

Analysis of single-nucleotide polymorphisms (SNPs) is important for diagnosis of infectious and genetic diseases, for environment and population studies, as well as in forensic applications. Herein is a detailed description to design an “operating cooperatively” (OC) sensor for ...

Hybridization-based techniques have been extensively employed for the analysis of specific DNA/RNA sequences. Herein, we describe highly specific inexpensive smart hybridization-based sensor that takes advantage of a universal molecular beacon probe as a fluorescent rep ...