遗传学实验技术

The Yeast Two-Hybrid (Y2H) system is the most frequently used method for identifying protein–protein interactions. The use of recombination-amenable Y2H vectors would reduce time and effort for cloning prey or bait vectors, and increase the quality of Y2H screenings due to the production ...

Full-length complementary DNAs (cDNAs) are an essential resource for functional genomics. Recently, we have developed a simple and efficient method for preparing a full-length cDNA library from a small amount of total RNA, named the “vector-capping” method. The biggest advantage of this ...

It is desirable to produce high homogeneity of novel fatty acids in oilseeds through genetic engineering to meet increasing demands by the oleo-chemical industry. However, expression of key enzymes for biosynthesis of industrial fatty acids usually results in low levels of desired fat ...

Unlike exponential amplification using polymerase chain reaction (PCR), linear RNA amplification using T7 RNA polymerase is advantageous for genome-wide analysis of gene expression and for cDNA library preparation from single-cell quantities of RNA. However, the use of RNA polym ...

Using yeast display, heterologous protein fragments can be efficiently displayed at high copy levels on the Saccharomyces cerevisiae cell wall. Yeast display can be used to screen large expressed protein libraries for proteins or protein fragments with specific binding propertie ...

We describe a novel expression cloning method based on screening yeast surface-displayed human cDNA libraries by direct affinity interaction to identify cellular proteins binding to a broad spectrum of target molecules. Being a eukaryote, yeast protein expression pathways are si ...

Single nucleotide polymorphisms (SNPs) are single base differences between haplotypes. SNPs are abundant in many species and valuable as markers for genetic map construction, modern molecular breeding programs, and quantitative genetic studies. SNPs are readily mined from gen ...

Transcriptome analysis by deep sequencing, more commonly known as RNA-seq is, becoming the method of choice for gene discovery and quantitative splicing detection. We published a double-random priming RNA-seq approach capable of generating strand-specific information . Poly(A)+   ...

Next-generation sequencing technologies have allowed new tools to be developed for transcriptome characterization. We describe here the methods for the preparation of a library from pooled RNAs for 454 sequencing of 3′-cDNA fragments. We also describe how the read sequences obtained ...

RNA interference (RNAi) using small interfering (siRNA) or short hairpin RNA (shRNA) has become the first choice for gene silencing maneuver in mammalian cells. Because different siRNAs of the same gene have variable silencing efficacy and only limited siRNAs are functional, many candi ...

Small RNAs (smRNAs) play an essential role in virtually every aspect of growth and development, by regulating gene expression at the post-transcriptional and/or transcriptional level. New high-throughput sequencing technology allows for a comprehensive coverage of smRNAs in any ...

Recent technological advances have enabled visualization of the organization and dynamics of local �chromatin structures; however, the global mechanisms by which chromatin organization modulates gene regulation are poorly understood. We designed and constructed a human a ...

DNA-based transposons are natural gene delivery vehicles. Similarly to retroviruses, these elements �integrate into the chromosomes of host cells, but their life-cycle does not involve reverse transcription and they are not infectious. Transposon-based gene delivery has seve ...

Mammalian artificial chromosomes (MACs) are engineered chromosomes with defined genetic content that can function as non-integrating vectors with large carrying capacity and stability. The large carrying capacity allows the engineering of MACs with multiple copies of the same ...

Advances in mammalian artificial chromosome technology have made chromosome-based vector technology amenable to a variety of biotechnology applications including cellular protein production, genomics, and animal transgenesis. A pivotal aspect of this technology is the a ...

The science of the so-called polymerase chain reaction (PCR) is now well known. However, the legal story associated with PCR is, for the most part, not understood and constantly changing. This presents diffculties for scientists, whether in academia or industry, who wish to practice the PCR proc ...

The development of the polymerase chain reaction (PCR) has often been likened to the development of the Internet, and although this does risk overstating the impact of PCR outside the scientific community, the comparison works well on a number of levels. Both inventions have emerged in the last 20 y ...

Polymerase chain reaction (PCR) is a very sensitive method of amplifying specific nucleic acid, but the system is susceptible to contamination from extraneous or previously amplified DNA strands (1,2). Many specific copies of DNA are produced from each round of amplification (3) with a sing ...

RNA extraction is fundamental to all aspects of mRNA analysis. We include here a simple method that avoids the use of a mortar and pestle.

Polymerase chain reaction (PCR), like any laboratory procedure, can be subject to a range of experimental or procedural error. A clear consideration of where such potential errors may occur is essential to minimize their impact. Careful quality control of equipment and reagents is essent ...