Ligation: purpose of the ligation reaction, plasmid DNA. plasmid vector, what is

Question

Ligation: purpose of the ligation reaction, plasmid DNA. plasmid vector, what is blunt-end ligation? What is a recombinant plasmid? Transformation: What arc competent cells? What is the purpose of introducing recombinant plasmid into bacterial cells? Why do you grow your transformed cells on ampicillin-containing agar plates? What is blue-white colony selection? How does it work? What is the advantage of blue and white selection? Plasmid preparation: What are the components is lysis buffer that breaks open bacterial cells. Why is alkali used in lysis buffer? Why bacterial genomic DNA and proteins precipitates out after neutralization of lysed cells with neutralization solution? how does the column purification of plasmid DNA works? Restriction digestion: What is a restriction enzyme? Why does your GAPC gene containing DNA insert fragment come out of the recombinant plasmid DNA after treating it with Bgl II restriction enzyme?

Explanation / Answer

Ligation:Blunt end ligation does not involve base-pairing of the protruding ends.Researchers can insert DNA fragments or genes into a plasmid vector, creating a so-called recombinant plasmid

Transformation:competence is the ability of a cell to take up extracellular DNA from its environment.purpose: cloning. antibiotic resistance selection for ampicilin.blue-white screen to eliminate false positive-blue,white positive. blue white screening works on X-gal method

Plasmid preparation:SDS and NaOH.SDS is there to solubilize the cell membrane. NaOH helps to break down the cell wall, but more importantly it disrupts the hydrogen bonding between the DNA bases, converting the double-stranded DNA (dsDNA) in the cell, including the genomic DNA (gDNA) and your plasmid, to single stranded DNA (ssDNA). This process is called denaturation and is central part of the procedure, which is why it

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