Table of Contents
What does pUC19 mean?
pUC19 is one of a series of plasmid cloning vectors created by Joachim Messing and co-workers. The designation “pUC” is derived from the classical “p” prefix (denoting “plasmid”) and the abbreviation for the University of California, where early work on the plasmid series had been conducted.
What is a plasmid Multicloning site?
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. Restriction sites within an MCS are typically unique, occurring only once within a given plasmid.
Why is pUC18 used?
Vectors pUC18 and pUC19 are small high-copy number plasmids that are widely used for cloning and manipulation of DNA fragments (9).
Is pUC19 artificial vector?
It is an artificial cloning vector created by Joachim Messing and his co-workers. It is a circular double-stranded DNA similar to the pBR322. But, pUC19 is widely used in molecular techniques due to its high copy number.
Is cloning site and recognition site same?
Cloning sites are the recognition sites of the restriction enzymes , it specific restriction enzyme site within a vector into which an insert was placed!
How do you read a plasmid map?
How to Read a Plasmid Map
- The name and size of the plasmid. The name and the size of the plasmid are indicated in the middle of the circular plasmid.
- The elements of a plasmid –
- The relative positions of the elements within the plasmid.
- The orientation of promoter.
What type of vector is pUC18?
Thermo Scientific pUC18 vector is a small, high copy number, E. coli plasmid, 2686 bp in length. It contains identical multiple cloning site (MCS) as pUC19 vector except that it is arranged in opposite orientation.
Where is pUC18 found?
pUC18 is isolated from E. coli strain HB101 by a standard plasmid purification procedure. Usage: pUC18 is a commonly used plasmid cloning vector in E. coli.
What is the purpose of pUC19 in the transformation?
pUC19 control DNA (10 pg/µl) is provided to check transformation efficiency. Use experimental DNA that is free of phenol, ethanol, salts, protein, and detergents to obtain maximum transformation efficiency.
Why is pUC19 a good cloning vector?
pUC19 is a commonly used cloning vector that conveys the Amp resistance. The molecule is a small double-stranded circle, 2686 base pairs in length, and has a high copy number.
What is pBR322 used for?
clone selection. Plasmid pBR322 is used extensively in genetic engineering. It has two genes of special interest. One codes for a protein that enables any host bacterium to resist the lethal effects of the antibiotic ampicillin and the other confers resistance to tetracycline.
Why does a vector need a single recognition site?
Answer: The vector requires very few or single recognition sites for the commonly used restriction enzymes. The presence of more than one recognition sites within the vector will generate several fragments leading to complication in gene cloning.
What does a plasmid map tell you?
Plasmid maps are graphical representation of plasmids, that show the locations of major identifiable landmarks on DNA like restriction enzyme sites, gene of interest, plasmid name and length etc.
Why are plasmid maps important?
Mapping of DNA restriction sites is an important part of working in a molecular biotechnology lab because such maps are used to plan cloning strategy and to verify when a DNA clone has been successfully constructed.
Why is pUC18 a good cloning vector?
pUC18 is a commonly used plasmid cloning vector in E. Due to a small size pUC18 enables successful cloning of large DNA fragments. The pUC18 plasmid confers ampicillin resistance and complement defects in β-galactosidase in appropriate host strains.
What is a good transformation efficiency for pUC19?
Transformation efficiency should be >1.0 × 1010 transformants/µg of pUC19 DNA.
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What is the cdcd spectrum of M13mp19 DNA?
CD spectra of M13mp19 DNA at 20°C as a function of NaCl concentration. The mean residue ellipticity with units of deg . cm2/decimole nucleotide is plotted on the y axis vs the wavelength, A, from 220 to 330 nm.
When was the M13mp19 circular single-strand DNA structure published?
Received August 10, 1988 Accepted February 10,1989 Title Structure and dynamics of M13mp19 circular single-strand DNA: Effects of ionic strength Created Date