提问
提问
我要登录
|免费注册
丁香实验推荐阅读
Generation of scFv from a Phage Display Mini-Library Derived from Tumor-Infiltrating B-Cells

B lymphocytes that infiltrate solid tumors, known as tumor-infiltrating lymphocyte B-cells (TIL-B-cells), are present in human tumors from different histological types (1,2) and tumor-specific antibodies can be detected in many cancer patients. For instance, anti-idiotypic an ...

丁香实验推荐阅读
Generation of Murine scFv Intrabodies from B-Cell Hybridomas

The intracellular expression of recombinant single-chain Fv (scFv) molecules—termed intrabodies—has potential advantages for therapeutic use (1,2). These single polypeptide chains are made of one heavy-chain variable region (VH) linked through a flexible spacer, usually a re ...

丁香实验推荐阅读
Quantitation of mRNA Levels by RT-PCR in Cells Purified by FACS: Application to Peripheral Cannabinoid Receptors in Leukocyte Subsets

Cell-sorting and mRNA amplification techniques with RT-PCR have been combined to study the gene expression of cytokines (1–3), receptors (4–6) and differentiation antigens of purified hematopoietic cells (7–9). Nevertheless, a major drawback of these techniques is the difficulty in ...

丁香实验推荐阅读
Detection of Anti-Sense RNA Transcripts by Anti-Sense RT-PCR

Reverse-transcriptase-polymerase chain reaction (RT-PCR) is a powerful and sensitive tool for RNA detection. This technique is particularly valuable in cases where the amount of an RNA species present is so minute that traditional RNA analysis methodologies such as Northern blot or R ...

丁香实验推荐阅读
Isolation of Coding Sequences from Genomic Regions Using Direct Selection

The rate-limiting steps in the positional cloning of disease genes are the identification of discrete genes within a large genomic region and the analysis of mutations in these genes. Direct selection is an expression-based gene identification technique that can rapidly identify cDN ...

丁香实验推荐阅读
Isolation of cDNAs Using the YAC Hybridization Screen Method

There are currently a number of techniques available for the isolation of novel coding sequences from cloned genomic fragments. The most recent methodologies include exon trapping (1), direct selection (2,3), and screening of cDNA libraries with whole radiolabeled yeast artificial c ...

丁香实验推荐阅读
Chemical Crosslinking Subtraction (CCLS)

Since the advent of polymerase chain reaction (PCR), it has proven possible to clone complete genes, or parts of these, without having to construct the usually obligatory cDNA library. PCR cloning, however, is not without its own problems and limitations, such as the fidelity of the cloned sequence ...

丁香实验推荐阅读
Detection and Isolation of Differentially Expressed Genes by Differential Display

Temporal and spatial expression of the 100,000 different genes in the genome of a mammal is a highly regulated process that determines the normal development of an organism. This assumption is based on the fact that only a fraction of these genes, in the range of 10,000–15,000, are expressed in any cell ty ...

丁香实验推荐阅读
Gene Mapping and Isolation: Access to Databases

If two variants in a population are related by simple Mendelian inheritance, then in principle, it is possible to isolate a gene in which the sequence differences will explain the nature of the variants. In practice, this is a daunting task in a genome the size of human (3000 Mb) and may take many years to achieve. ...

丁香实验推荐阅读
Software for Genetic Linkage Analysis: An Update

Reviews describing the software available to support genetic linkage analysis have appeared intermittently over the last few years (1,1a). This period has been associated with several significant influences that have made it timely to reconsider the available programs and related ...

丁香实验推荐阅读
Gene Mapping Goes from FISH to Surfing the Net

The arrival of the second millennium will usher in unsurpassed information and knowledge of our genetic constitution, and will promise to revolutionize basic research and molecular medicine. The road toward a complete understanding of our genetic makeup is largely the fruit of the Human G ...

丁香实验推荐阅读
Exon Detection by Similarity Searches

Other chapters of this volume have presented the various experimental methods (mainly exon trapping and recombination-based and hybridization-based approaches) used for the identification of transcribed sequences within cloned genomic fragments. None of those methods re ...

丁香实验推荐阅读
Linkage Analysis of Genetic Disorders

Genetic disorders follow a classic Mendelian dominant or recessive single-locus pattern of inheritance or a complex genetic pattern (multiple genes and environmental influences). In general, the complexity arises when the simple correspondence between genotype and phenoty ...

丁香实验推荐阅读
Gene Ordering and Localization by Linkage Analysis

The essence of linkage analysis is a deviation from the Mendelian principle of independent, or random, assortment of gene pairs when transmitted from generation to generation. Two genes are said to be completely linked (see Section 4.1. for definitions of genetic terms) when there is no recombi ...

丁香实验推荐阅读
Gene Mapping Using Somatic Cell Hybrids

Human-rodent somatic cell hybrids have proven to be a useful tool for mapping expressed gene products (1,2) and, in particular, DNA sequences. The accuracy of the method is dependent on the identification of a species difference between the human and rodent species. This chapter details metho ...

丁香实验推荐阅读
Probe Ordering and Distancing by FISH

In situ hybridization (ISH) has proven to be a powerful methodological approach to visualize specific nucleic acid sequences directly within the morphological context of the cell. The method involves hybridization of labeled probes to denatured target chromatin that has been fixed on ...

丁香实验推荐阅读
Physical Mapping by Pulsed-Field Gel Electrophoresis

During the past few years, there has been intense activity in physically mapping the genomes of a variety of organisms, including human, mouse, Drosophila, Caenorhabditis elegans, Arabidopsis thaliana, and yeast. The considerable advances made in physically mapping these genomes has ...

丁香实验推荐阅读
Construction and Use of Cosmid Contigs

The application of cosmids to positional cloning strategies has changed over the last several years (1). Initially, the construction of genome physical maps, for example, in Caenorhadbitis elegans and Arabidopsis (2,3), was attempted using cosmid clones randomly selected and analyz ...

丁香实验推荐阅读
Construction and Use of YAC Contigs in Disease Regions

The identification of genes by positional cloning has revolutionized the molecular analysis of disease. Disease-causing genes can now be isolated with no knowledge of the biological function of the molecule involved, relying only on genetic inheritance or chromosomal perturbat ...

丁香实验推荐阅读
Reduced Representation Methods for Subgenomic Enrichment and Next-Generation Sequencing

Several methods have been developed to enrich DNA for subsets of the genome prior to next-generation sequencing. These front-end enrichment strategies provide powerful and cost-effective tools for researchers interested in collecting large-scale genomic sequence data. In this ...

提问
扫一扫
丁香实验小程序二维码
实验小助手
丁香实验公众号二维码
关注公众号
反馈
TOP
打开小程序