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    Menna Elbaroudy
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    Polymerase chain reaction (PCR) enables large amounts of DNA to be produced from very small samples through repeating cycles of separating and synthesizing DNA strands. The process involves placing a DNA sample, nucleotides, DNA primers, and a thermostable DNA polymerase in a PCR machine. The strands of DNA are then separated by heating and cooled to allow the primers to bind. The mixture is then heated again for replication by the polymerase. This cycle repeats many times, resulting in exponential amplification of the target DNA region flanked by the primers. PCR requires a thermostable polymerase and pure nucleotides and DNA to avoid contamination. It has many uses including in legal cases but is currently suspended in the UK.

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    PCR

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    Polymerase chain reaction (PCR) enables large amounts of DNA to be produced from very small samples through repeating cycles of separating and synthesizing DNA strands. The process involves placing a DNA sample, nucleotides, DNA primers, and a thermostable DNA polymerase in a PCR machine. The strands of DNA are then separated by heating and cooled to allow the primers to bind. The mixture is then heated again for replication by the polymerase. This cycle repeats many times, resulting in exponential amplification of the target DNA region flanked by the primers. PCR requires a thermostable polymerase and pure nucleotides and DNA to avoid contamination. It has many uses including in legal cases but is currently suspended in the UK.

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    23 views14 pages

    Understanding PCR Process and Its Uses

    Uploaded by

    Menna Elbaroudy
    Polymerase chain reaction (PCR) enables large amounts of DNA to be produced from very small samples through repeating cycles of separating and synthesizing DNA strands. The process involves placing a DNA sample, nucleotides, DNA primers, and a thermostable DNA polymerase in a PCR machine. The strands of DNA are then separated by heating and cooled to allow the primers to bind. The mixture is then heated again for replication by the polymerase. This cycle repeats many times, resulting in exponential amplification of the target DNA region flanked by the primers. PCR requires a thermostable polymerase and pure nucleotides and DNA to avoid contamination. It has many uses including in legal cases but is currently suspended in the UK.

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    of 14

    18 March 2012

    Polymerase Chain Reaction


    Aims To understand the process of PCR and its uses.
    Starter - Match each term with its correct description (work in pairs)

    What is it?

    Animations
    http://www.dnalc.org/ddnalc/resource s/pcr.html http://www.maxanim.com/genetics/PC R/pcr.swf

    Information
    Polymerase

    chain reaction enables large amounts of DNA to be produced from very small samples (0.1ml) There is a repeating cycle of: separation of double DNA strands synthesis of a complementary strand for each

    What happens?
    Sample DNA , nucleotides, DNA primers & thermostable DNA polymerase placed in PCR machine. Strands of sample DNA separated by heating to 95oC Mixture cooled to 37oC to allow primers to bind. Mixture heated to 72oC for replication (optimum temp of DNA polymerase) Cycle repeats many times (~8mins /cycle)

    Problems
    Separation achieved by heating to 95oC no suitable helicase DNA polymerase cant work on completely single stranded DNA double stranded regions needed at the start of sequence to be copied: primers (short sequences DNA) complementary to bases at start of region to be copied used To synthesize primers , base sequence at start must be known

    TTAACGGGGCCCTTTAAA.....TTTAAACCCGGGTTT
    AATTGCCCCGGGAAATTT.....AAATTTGGGCCCAAA
    the

    sequence of bases which ONLY flank a particular region of a particular organism's DNA, and NO OTHER ORGANISM'S DNA. This region would be a target sequence for PCR.

    The first step for PCR would be to synthesize "primers" of about 20 letterslong ONE primer exactly like the lower lefthand sequence, and ONE primer exactly like the upper right-hand sequence:

    TTAACGGGGCCCTTTAAA.....TTTAAACCCGGGTT AATTGCCCCGGGAAATTT.......................>

    and:
    <........................................TTTAAACCCGGGTTT AATTGCCCCGGGAAATTT........AAATTTGGGCCCAAA

    DNA

    polymerase must be thermostable (37oC 95oC used) to avoid fresh enzyme being added.

    Remember.
    Nucleotides

    used must be very pure. DNA must not be contaminated any foreign DNA would also be copied.PCR in legal cases in UK suspended at present

    Uses of PCR
    http://nobelprize.org/nobel_prizes/che mistry/laureates/1993/illpres/already .html

    This powerpoint was kindly donated to www.worldofteaching.com

    http://www.worldofteaching.com is home to over a thousand powerpoints submitted by teachers. This is a completely free site and requires no registration. Please visit and I hope it will help in your teaching.

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