Open AccessJournal Article
New primer strategy improves precision of differential display
TLDR
Modified long composite primers were developed based on both mRNA differential display and RNA arbitrarily primed PCR fingerprinting methods to increase the reproducibility and sensitivity of the mRNA differentialdisplay while still keeping the characteristics of the original method.Abstract:
To increase the reproducibility and to reduce the false positives in the initial mRNA differential display, modified long composite primers were developed based on both mRNA differential display and RNA arbitrarily primed PCR fingerprinting methods. Ten-base nucleotides were added at the 5' ends of the primers used in the initial mRNA differential display. These included a restriction site to aid cloning. PCR began with one low-stringency cycle (40 degrees C for annealing) followed by 35 high-stringency cycles (60 degrees C for annealing). The modified method significantly improved the reproducibility and sensitivity of the mRNA differential display while still keeping the characteristics of the original method.read more
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Book ChapterDOI
Principles of differential display.
TL;DR: This chapter describes the principles of DD, including specific methods for working with the primers that have been used extensively, and discusses the production of a DD gel, strategies for isolating and identifying DD band cDNAs, and information on performing high-throughput DD.
Journal ArticleDOI
Applications of Differential-Display Reverse Transcription-PCR to Molecular Pathogenesis and Medical Mycology
TL;DR: DDRT-PCR studies identified TIF-2, which may play a role in the upregulation of phospholipases, and the stress-related genes, CIP1 and CIP2, and genes involved in the host-pathogen interaction, including a member of the 100-kDa family in Histoplasma and an ALS and 14-3-3 gene in Candida, were potentially identified by the procedure.
Journal ArticleDOI
Retroviral-mediated transmission of a mouse VL30 RNA to human melanoma cells promotes metastasis in an immunodeficient mouse model
TL;DR: Southern hybridization analyses showed that the mVL30-1 cDNA integrated into different genomic sites in different melanoma clones, suggesting that the effect of the m Vlad 30 (mVL30) retroelement on metastasis depends not on integration per se but instead on expression of themVL 30-1 RNA.
Journal ArticleDOI
Identification of cystatin B in human esophageal carcinoma, using differential displays in which the gene expression is related to lymph-node metastasis
TL;DR: This study suggests that reduced expression of cystatin B in esophageal‐carcinoma tissue is associated with lymph‐node metastasis and may therefore prove to be a useful marker for predicting the biologic aggressiveness of human esophagal carcinoma.
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