Detection of eukaryotic cDNA in differential display is enhanced by the addition of E. coli RNA.
نویسندگان
چکیده
We describe a method to enhance the sensitivity of eukaryotic cDNA detection in differential display (DD). Typically, DD protocols require between 200 and 500 ng RNA for each reverse transcription reaction. The addition of Escherichia coli RNA before reverse transcription of eukaryotic RNA increases the detection of DD patterns more than tenfold. The method broadens the applicability of DD and allows the identification of genes that are differentially expressed when the amount of eukaryotic RNA is limited.
منابع مشابه
Enhanced Bioadsorption of Cadmium and Nickel by E. coli Displaying A Metal Binding Motif Using CS3 Fimbriae
Display of peptides on the surface of bacteria offers many new and exciting applications in biotechnology. Fimbriae is a good candidate for epitope display on the surface of bacteria. The potential of CS3 fimbriae of enterotoxigenic E. coli as a display system has been investigated. A novel cell surface display system with metal binding property was developed by using CS3 fimbriae. Short metal ...
متن کاملDifferential Expression of Arabidopsis thaliana Acid Phosphatases in Response to Abiotic Stresses
The objective of this research is to identify Arabidopsis thaliana genes encoding acid phosphatases induced by phosphate starvation. Multiple alignments of eukaryotic acid phosphatase amino acid sequences led to the classification of these proteins into four groups including purple acid phosphatases (PAPs). Specific primers were degenerated and designed based on conserved sequences of PAPs isol...
متن کاملDetection of Viable But Non-Culturable State of Escherichia coli O157:H7 Using Reverse Transcription PCR
Background and Aims: Many bacteria including Escherichia coli may enter into a viable but non-culturable (VBNC) state under unfavorable stresses, which are unable to be detected by culture-based methods. In this study, the use of Reverse Transcription PCR (RT-PCR) for detection of VBNC state of E. coli O157:H7 was investigated. Materials and Methods: Escherichia. coli O157:H7 was inoculated i...
متن کاملCloning and Expression of Coxsakievirus B3 Viral Protein-1 in E. Coli
Viral protein-1 (VP1) is a major capsid protein of Coxsakievirus B3 (CVB3) that plays an important role in directing viruses towards permissive cells and acts as a main antigenic site of the virus in eliciting of host immune response, hence it seems VP1 can be considered as a vaccine candidate against CVB3 infection. In this study, cDNA of VP1 was prepared, cloned into pET expression vector and...
متن کاملEffects of ackA, pta and poxB inhibition by antisense RNA on acetate excretion and recombinant beta interferon expression in Escherichia coli
Introduction: Escherichia coli (E.coli) is one of the most widely used hosts for the production of recombinant proteins. The main problem in getting high product yields and productivity is the accumulation of acetic acid (acetate) as an unwanted metabolic by-product. In this study, an antisense-based strategy as a metabolic engineering approach was employed to hamper the acetate excretion probl...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
- BioTechniques
دوره 28 1 شماره
صفحات -
تاریخ انتشار 2000