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A transformation efficiency of 1×10 8 cfu/μg for a small plasmid like pUC19 is roughly equivalent to 1 in 2000 molecules of the plasmid used being introduced into cells. In E. coli , the theoretical limit of transformation efficiency for most commonly used plasmids would be over 1×10 11 cfu/μg.
A pUC19 cloning vector showing the multiple cloning site sequence with restriction enzyme sites. A multiple cloning site (MCS), also called a polylinker, is a short segment of DNA which contains many (up to ~20) restriction sites—a standard feature of engineered plasmids. [1]
Cells which have been successfully transformed with pUC19 can be differentiated from cells which have not by growing them on media with ampicillin. Only the cells with the plasmid containing amp R will survive. The origin of replication (ori), is derived from the plasmid pMB1. [6] [1] pUC19 is a high copy number plasmid. [3]
They are the standard cloning vectors and the ones most commonly used. Most general plasmids may be used to clone DNA inserts of up to 15 kb in size. One of the earliest commonly used cloning vectors is the pBR322 plasmid. Other cloning vectors include the pUC series of plasmids, and a large number of different cloning plasmid vectors are ...
Sequence coverage (or depth) is the number of unique reads that include a given nucleotide in the reconstructed sequence. [1] [2] Deep sequencing refers to the general concept of aiming for high number of unique reads of each region of a sequence. [3] Physical coverage, the cumulative length of reads or read pairs expressed as a multiple of ...
[1] One approach in constructing a restriction map of a DNA molecule is to sequence the whole molecule and to run the sequence through a computer program that will find the recognition sites that are present for every restriction enzyme known. Before sequencing was automated, it would have been prohibitively expensive to sequence an entire DNA ...
(Typical transgene delivery methods involve plasmids, which contain foreign DNA.) The smaller size of minicircles also extends their cloning capacity and facilitates their delivery into cells. Their preparation usually follows a two-step procedure: [4] [5] production of a 'parental plasmid' (bacterial plasmid with eukaryotic inserts) in E. coli
P1 vectors also contain a P1 plasmid replicon, which ensures only one copy of the vector is present in a cell. However, there is a second P1 replicon- called the P1 lytic replicon- that is controlled by an inducible promoter. This promoter allows the amplification of more than one copy of the vector per cell prior to DNA extraction. [2] bac vector