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    Primer

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    Iqra fiaz

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    © All Rights Reserved
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    Primer

    Uploaded by

    Iqra fiaz
    2 views6 pages

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    primer

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    Copyright:

    © All Rights Reserved
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    Download as pptx, pdf, or txt
    2 views6 pages

    Primer

    Uploaded by

    Iqra fiaz

    Copyright:

    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
    of 6

    Primer Designing for PCR

    Polymerase chain reaction (abbreviated PCR) is a laboratory


    technique for rapidly producing (amplifying) millions to billions of
    copies of a specific segment of DNA, which can then be studied in
    greater detail.
     PCR involves using short synthetic DNA fragments called primers to
    select a segment of the genome to be amplified, and then multiple
    rounds of DNA synthesis to amplify that segment.
    • To amplify any DNA sequence, two primers are necessary. One is
    called ‘forward primer' and the other one is called ‘reverse primer’.
    The forward primer synthesizes the upper strand using the bottom
    strand as a template. Whereas Reverse primer uses the upper strand
    as a template and synthesizes the lower strand.
    Specification of primer
    • Length of 18-24 bases
    • 40-60% G/C content
    • Start and end with 1-2 G/C pairs
    • Melting temperature (Tm) of 50-60°C
    • Primer pairs should have a Tm within 5°C of each other
    • Primer pairs should not have complementary regions
    • Annealing temperature of a primer should be lower than Tm
    • Avoid runs of 4 or more of one base, or dinucleotide repeats (for
    example, ACCCC or ATATATAT
    Tools for primer Designing
    • Primer Blast
    • Primer 3
    • Oligo calculator

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