Nonisotopic probes have been widely adopted for DNA fingerprinting and DNA profiling because of their ease and speed of use and obvious safety and environmental advantages (1).
The possibility of amplifying total complementary DNA (cDNA) obtained from small amounts of biological material is not yet routinely considered, despite the fact that obtaining amounts of material suitable for direct processing by standard methods is often time-consuming and exp ...
Several existing methods for constructing complementary DNA (cDNA) libraries require several thousand cells and involve the lengthy procedure of reverse transcription (RT), restriction, adaptor ligation, and vector cloning, which usually fail to maintain the completeness of a ...
Molecular profiling of single-cell gene expression permits the high-definition investigation of intracellular gene activity and physiological status of the cells under certain special conditions, such as pathogenesis (1,2), cancer staging (3), drug treatment, and developm ...
A perfect complementary DNA (cDNA) library would provide an accurate and complete representation of all messenger RNA (mRNA) sequences expressed in a particular source, whether a cell, tissue, or organism. Of primary importance during cDNA library construction is the use of a high-quali ...
Gel electrophoresis has been widely used in separating and purifying macromolecules such as proteins and nucleic acids that differ in size, shape, charge, and conformations. The gel basically acts as a tube in which the long-chain macromolecules such as RNAs contract and migrate in an extend ...
The generation and measurement of low-abundance messenger RNA (mRNA) transcripts is possible using techniques such as reverse transcriptas-polymerase chain reaction (RT-PCR) (1,2). There are a number of variables that must be controlled for if accurate and reproducible results are ...
The debut of RNA-polymerase cycling reaction (RNA-PCR) has promised to provide linear amplification of a reproducible mRNA library from as few as 20 single cells (1). By incorporating a RNA promoter element during the synthesis of double-stranded complementary DNA (cDNA) templates, a pol ...
Complementary DNA (cDNA) library construction is a foundational technique for various organism genome projects and molecular biological approaches. Numerous methods for generating a cDNA library have been applied for different purposes. To profile the gene expression of a given ...
The goal for developing the SPGI (screening poly cDNA for gene identification technique was to generate an efficient tool for maximal identification of the expressed genes from eukaryotic genomes. The impetus for developing the SPGI method was triggered by the observation that a large nu ...
Serial analysis of gene expression (SAGE) is a powerful technique for genomewide analysis of gene expression (1–14). However, almost two-thirds of SAGE tags cannot be used directly for gene identification for two reasons. First, many of SAGE tags match to multiple known expressed sequences b ...
Complementary DNA libraries reflect gene expression at certain times for specific cells, whereas genomic DNA libraries represent all genetic information in somatic cells. The complexity of cellular organization reflects a genetic program that encodes a collection of genes and the ...
With sequence analysis of the human genome well underway, the scientific focus is shifting toward understanding the fundamentals of gene function. Sequence information alone is insufficient for a full understanding of gene function, expression, and regulation. Modern approach ...
Subtractive hybridization is a technique for detecting differences of gene expression between two populations of RNAs or complementary DNAs (cDNAs). It is based on base pair complementary that nucleic acid sequences in common with the two populations can form hybrids. After hybridiz ...
The ability to compare two different messenger RNA (mRNA) or complementary DNA (cDNA) libraries has permitted studies into the role of differentially expressed genes involved in the mechanisms of neoplastic transformation, developmental regulation, therapeutic effects, pa ...
The polymerase chain reaction (PCR) is a powerful method permitting generation of almost any desired cDNA sequence. Construction of cDNA fragments that encode repetitive amino acid sequences by PCR has proven problematic, however, as repetitive nucleotide primers tend to amplify un ...
The generation of peptide from messenger RNA (mRNA) provides a convenient source for current proteomic analysis. Intron-free mRNA possessing adenine-uracil-guanine (AUG) start codons can be translated into labeled or unlabeled peptides under a predetermined reticulocyte ly ...
Complementary DNA (cDNA) libraries represent the information encoded in the messenger RNA (mRNA) of a particular tissue or organism. RNA molecules are exceptionally labile and difficult to amplify in their natural form. For this reason, a cDNA library is created by converting the informa ...
The BacterioMatch™ two-hybrid system provides a fast, simple, and efficient method for in vivo detection of protein-protein interactions in Escherichia coli (1,2). This system is based on transcriptional activation of a reporter gene. In this system, a protein of interest (the bait) is fused ...
We have made tremendous strides in our understanding and utilization of the new methodologies in molecular biology over the past 50 yr. With our recent access to complete genome sequences, the function of different transcripts and proteins, the gene products, will soon be determined and, con ...
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