Plasmid sequences

Whereas almost any DNA sequence is a possible tool for IPCR, interestingly, a large number of publications described the usage of a standard plasmid sequence from pBluescript/M13 [25, 27, 33-36] or pUC19 [10, 13, 26, 28, 37] as a kind of standard IPCR marker derived from DNA ready available in the typical molecular biology laboratory. Maia et al. emphasize the importance of a DNA-marker not present in the routine work of the laboratory carrying out IPCR to avoid cross-contamination. In their work, they used a unique BTV-virus-DNA sequence only found in ruminants for the detection of HbsAg [30],... 

Input a desired plasmid sequence via File -> Open. 

Use as a tool (see Note 1) for genome search and annotation that integrates microbial genome sequence data with most eukaryotic sequence data as well as viral and plasmid sequences (8). 

Materials pJK502 is a T7-overexpression vector that encodes for wild-type E.coli RNase HI. It was made by site-directed mutagenesis of pSMlOl (24), reverting three alanines (residues 13, 63 and 133) to cysteines, and then subcloning the resulting gene into a pETlla overexpression vector (details of plasmid sequence available upon request). Overexpression and purification of E.coli RNase HI were performed essentially as described in (24) for RNase H. RNase H is... 

Once constructed, the plasmid sequences are introduced into the virus chromosome by co-transfection of the plasmid with naked wild type viral DNA. Recombination occurs at a low but useful frequency between these DNAs at their homologous regions, flanking the polyhedrin gene. The progeny virus from the co-transfection are plaqued, and the plaques are scored for type. Under a dissecting microscope, the recombinants can be selected from the wild types based on their distinctive plaque morphology due to a lack of PIBs. Once selected and a stock prepared, one has a virus which on infection of cells will direct the production of both additional virus and the product of the... 

Inc rude plasmid sequence p1 Select sequences from single genus p... 

Plasmid (sequencer) ThermoFidelase PGEM (ABI) pBSC (ABI) PSPC (ABI) pUC (ALF)... 

If one cannot include I-Scel in the strategy for making a transgenic, the DNA should be linearized or the insert should be isolated from the plasmid sequences. Supercoiled DNA, however, can be used for transient gene expression work. After phenol-chloroform extraction to remove the restriction enzyme, the DNA is further purified on a MicroSpin column (MicroSpin S-300HR, Amersham Biosciences, Piscataway, NJ) as follows ... 

Jumping clone arms can be separated from the plasmid sequences using either the enzyme of the first digestion step (Notl, Mlul), or, if this site has been destroyed (e.g., by the use of an Mlul cleaved plasmid to construct a BssHII jumping library), by flanking sites from the polylinker (see Table 2). 

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