Methods used encountered li transformation, pcr, restriction enzymes, plasmid. Isolation, restriction digestion, and gel electrophoresis of plasmid dna prathyusha gudapati, biol 304, spring 2015. Mobilization and prevalence of a fusobacterial plasmid. Home resources plasmid files basic cloning vectors pbluescript ks pbluescript ks standard cloning vector phagemid excised from lambda zap. The pbluescript ii phagemids plasmids with a phage origin are cloning vectors designed to simplify commonly used cloning and sequencing procedures, including the construction of nested deletions for dna sequencing, generation of rna transcripts in vitroand sitespecific mutagenesis and gene mapping. To provide the tool with more advantages for molecular cloning in e. Featuring an extensive polylinker with 21 unique restriction enzyme recognition sites, the vectors are suitable for. Agilent pbluescript ii vectors are powerful cloning vectors for a range of research applications. Abstract the ezh2 enhance of zeste homolog2 is an enzyme that in humans is. In order to further investigate its effect on dna, it was recloned into pbluescript plasmid with histag. Agilent technologies pbluescript ii phagemid vectors.
The ampicillinresistance gene has been replaced with the chloramphenicol resistance gene. Isolation, restriction digestion, and electrophoresis of. This is a free resource for the scientific community that is compiled by addgene this page is informational only this vector is not available from addgene please contact the manufacturer for further details. Problems with transformation pbluescript plasmid dna i have a gene, apoe, and transformed it into pbluescript digested with ecorv with suitable antibiotic in the right concentration. The vectors contain polylinkers that are designed to optimize a number of. Click any vector name to download the vector map in pdf. Restriction mapping restriction mapping is a method used to map an unknown segment of dna by breaking it into pieces and then identifying the locations of the breakpoints. The bluntend pcr products were cloned in plasmid pbluescript ii sk altingmees and.
Phagemid vectors a phagemid or phasmid is a type of cloning vector developed as a hybrid of the filamentous phage m and plasmids to produce a vector that can grow as a plasmid, and also be. Abstract the purpose of the experiment was to isolate plasmid dna, followed by. Sequence extractor generates a clickable restriction map and pcr primer map of. Cloning of plasmid ezh2 gene into pbluescript ii ks. Broad host rangeli plasmid vector pghost4 complete broad host rangeli plasmid vector pghost5 complete broad host rangeli plasmid. Cosmid vectors for rapid genomic walking, restriction mapping, and gene transfer.
Wahl gm, lewis ka, ruiz jc, rothenberg b, zhao j, evans ga. Phagemid vectors biology read, write, learn and apply. Help pages, faqs, uniprotkb manual, documents, news archive and biocuration projects. The ampicillinresistanceencoding gene ap r of pbluescript has been replaced by genes encoding resistance to either kanamycin km r or tetracycline tc r. Get readytouse clones in the pet vector or other expression vector of your choice.
Welcome to vector database vector database is a digital collection of vector backbones assembled from publications and commercially available sources. We notice that you are using internet explorer, which is not supported by xenbase and may cause the site to display incorrectly. It is a cloned carrier, which is derived from phage. Standard cloning vector phagemid excised from lambda zap. The video give a basic introduction to plasmid vectors. In vitro rna transcription f1 origin in orientation, kpnsac polylinker orientation, contains. Phagemids represent a hybrid type of class of vectors that serve to produce singlestranded dna. A new set of useful cloning and expression vectors derived. The construction and purification of recombinant baculovirus vectors for the expression of foreign genes in insect cells by standard transfection and plaque assay methods can take as long as 4 to 6 weeks.
The pbluescript ii sk and ks vector series represent two orientations of the mcs multiple cloning site within the lacz gene encoding the nterminal fragment of betagalactosidase ks. When your child keeps you up all night long and you cant get more than two or three hours of shuteye at a time, it. The bluntend pcr products were cloned in plasmid pbluescript ii sk alting mees and short, 1989 digested with ecorv, and their dna sequence was. The pbluescript ii phagemids plasmids with a phage origin are cloning vectors designed to simplify commonly used cloning and sequencing procedures, including the construction of nested deletions for. Problems with transformation pbluescript plasmid dna. Phagemid cloning system t7 and t3 promoters m forward and m reverse sequencing primers ampicillin resistance in bacterial cultures. This tutorial shows you how to choose and map the correct pbluescript plasmid either ks or sk, for you to use for making your recombinant plasmid. About phagemid before introducing the phagemid, it is important for us to know something related to itthe plasmid. Theres a reason its commonly used as a form of torture. What is a phagemid vector this article introduces some basic information of phagemid, and phagemid vector, as well as the related biotech companycreative biolabs.
The origin of dna replication ori, conferring to pbluescript. Ezh2 protein altered chromatin structure by methylation making it inaccessible to dna polymerase. How can i be notified when a plasmid from a specific lab or paper is available. We suggest using a current version of chrome, firefox, or safari. This vector is often used in cloning and sequencing, including dna sequencing, nested deletion, rna transcription, site directed mutagenesis and gene mapping. The plasmid vectors, pbs2ndd and pbs3ndd, for versatile cloning. This vector is often used in cloning and sequencing, including dna sequencing, nested deletion, rna transcription, site directed mutagenesis and gene mapping in vitro.