Tom asked in Science & MathematicsBiology · 1 decade ago

Forward and Reverse Primers in PCR?

How do you design forward and reverse primers? My textbook and lecture notes give me nothing to work with. Lets say the region that you want to copy is-

5' ...ACGAACTGGCGTAGGATCGTAAGTCAGT... 3'

3' ...TGCTTGACCGCATCCTAGCATTCAGTCA... 5'

What would be an appropriate forward and reverse primer and why? Anything would be much appreciated!

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  • 1 decade ago
    Favorite Answer

    This is a very short region to amplify, so let`s say instead that it`s:

    5' ...ACGAACTGGCGTA ... GGATCGTAAGTCAGT... 3'

    3' ...TGCTTGACCGCAT ... CCTAGCATTCAGTCA... 5'

    This is the same sequence, with the amplified region indicated by the dots in the centre. Primers are usually 10-20 bp long, so these might be one the short side, but your primers would be:

    5' ACGAACTGGCGTA 3` as your `forward primer`

    and 3`CCTAGCATTCAGTCA 5' as your `reverse primer`

    Remember that DNA polymerase needs a 3` OH to add to, so the primer anneals to the DNA and is extended from the 3` end. The 3` ends of these primers point into the region you want to amplify.

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  • hocker
    Lv 4
    3 years ago

    Forward And Reverse Primers

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  • Anonymous
    5 years ago

    This Site Might Help You.

    RE:

    Forward and Reverse Primers in PCR?

    How do you design forward and reverse primers? My textbook and lecture notes give me nothing to work with. Lets say the region that you want to copy is-

    5' ...ACGAACTGGCGTAGGATCGTAAGTCAGT... 3'

    3' ...TGCTTGACCGCATCCTAGCATTCAGTCA... 5'

    What would be an appropriate forward and...

    Source(s): reverse primers pcr: https://biturl.im/gdVNt
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  • Anonymous
    6 years ago

    Forward and reverse, sense and antisense primers:

    http://www.youtube.com/watch?v=0vdbf4F90_o

    Youtube thumbnail

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  • 5 years ago

    to separate the two DNA strands (the ladder) i.e. the separation of the ladder to make more DNA ladders each copying the other side of the ladder (forward and reverse)

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  • 4 years ago

    They are not complementary to each other.

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  • Anonymous
    5 years ago

    IBD2 F TCACCGTCCTCAGCTTAC

    IBD2 R TCAGGATTTGGGATCAGC

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  • 6 years ago

    please tell me why we use high temperature in pcr

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