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Library Creation – Obtain DNA sample, Break DNA into pieces

2 comments to Library Creation – Obtain DNA sample, Break DNA into pieces

  • yusuf

    -a gene library is a set of thousands of DNA fragments from one organism

    - DNA of organism is cut using a restriction endonuclease

    - each fragment is inserted into plasmid to make a recombinant plasmid

    -recombinant plasmids are then inserted in bacterial cells

    -mRNA is isolated from cells

    -reverse transcriptase is used to make a DNA copy of the mRNA

    -use DNA polymerase to make the single stranded cDNA double stranded

    - each piece of DNA is added to a seperate plasmid and inserted into a bacterial cell

  • Mohamed Ahmed

    Too complicated, the book is much easier just 3 steps:
    1-isolate mrna from cells.
    2-add reverse transcriptase to make single strand cdna, add dna polymerase to make it double stranded.
    3-add dna to plasmid and insert in e.coli cells.

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