Direct PCR Screening of Pichia pastorisClones
نویسندگان
چکیده
منابع مشابه
Random PCR mutagenesis screening of secreted proteins by direct expression in mammalian cells.
We have developed a general method for screening randomly mutagenized expression libraries in mammalian cells by using fluorescence-activated cell sorting (FACS). The cDNA sequence of a secreted protein is randomly mutagenized by PCR under conditions of reduced Taq polymerase fidelity. The mutated DNA is inserted into an expression vector encoding the membrane glycophospholipid anchor sequence ...
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Researchers often purify DNA from blood samples prior to performing PCR because it is believed that blood constituents and the reagents commonly used to preserve blood samples (e.g., anticoagulants) interfere with PCR. In order to save the time and expense required for template purification, several methods have been reported for direct PCR of blood samples. These include microwave irradiation,...
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genes. Methods Enzymol. 185:60-89. 5.Switzer, R.C., C.R. Merril and S. Shifrin. 1979. A highly sensitive silver stain for detecting proteins and peptides in polyacrylamide gels. Anal. Biochem. 98:231-237. 6.Tabor, S. and C.C. Richardson. 1985. A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes. Proc. Natl. Acad. Sci. USA 82:1074-1078. 7.Whela...
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Successful DNA isolation is a critical step in molecular studies of symbiotic fungi including plant pathogens, mycorrhizal fungi, endophytic fungi and lichen-forming fungi. Whereas standard isolation protocols are usually the straightforward approach if axenic cultures are available, it is problematic to isolate DNA from fungi that grow in intricate associations with some other organisms. If th...
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ژورنال
عنوان ژورنال: BioTechniques
سال: 1996
ISSN: 0736-6205,1940-9818
DOI: 10.2144/96206bm08