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  • Calculations For PCR

    Încărcat de

    Irazema Fuentes
    40 vizualizări6 pagini
    The document provides reaction mix formulations for PCR amplifications of β-tubulin, A-Elongation Factor, MAT1, and MAT2. It lists the components, volumes, and stock concentrations needed to make reaction mixes ranging from 1 to 100 reactions, each with a total volume of 20, 20, 50, or 50 μl respectively.

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    Titlu original

    Calculations for PCR

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    © Attribution Non-Commercial (BY-NC)

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    The document provides reaction mix formulations for PCR amplifications of β-tubulin, A-Elongation Factor, MAT1, and MAT2. It lists the components, volumes, and stock concentrations needed to make reaction mixes ranging from 1 to 100 reactions, each with a total volume of 20, 20, 50, or 50 μl respectively.

    Drepturi de autor:

    Attribution Non-Commercial (BY-NC)
    Descărcați ca XLS, PDF, TXT sau citiți online pe Scribd
    Indicator pentru conținut neadecvat
    40 vizualizări6 pagini

    Calculations For PCR

    Încărcat de

    Irazema Fuentes
    The document provides reaction mix formulations for PCR amplifications of β-tubulin, A-Elongation Factor, MAT1, and MAT2. It lists the components, volumes, and stock concentrations needed to make reaction mixes ranging from 1 to 100 reactions, each with a total volume of 20, 20, 50, or 50 μl respectively.

    Drepturi de autor:

    Attribution Non-Commercial (BY-NC)
    Descărcați ca XLS, PDF, TXT sau citiți online pe Scribd
    Indicator pentru conținut neadecvat
    din 6

    Reaction mixes for β-tubulin amplifications (20-

    μl reaction volumes)

    1x 5 10 20 30 40

    Target DNA (1-20 ng/μl) 1 add separately

    10× PCR Buffer 2 10 20 40 60 80

    10× Mg++ (25 mM) stock 1.2 6 12 24 36 48

    10× dNTPs (2 mM) stock 2 10 20 40 60 80

    10× T1 primer stock (2.5


    2 10 20 40 60 80
    pmoles/μl) stock
    10× T2 primer stock (2.5
    2 10 20 40 60 80
    pmoles/μl) stock

    Taq DNA polymerase 0.1 0.5 1 2 3 4

    Double-distilled H2O 9.7 48.5 97 194 291 388

    total volume 20

    Reaction mixes for A-Elongation Factor


    amplifications (20-μl reaction volumes)

    1x 5 10 20 30 40

    Target DNA (1-20 ng/μl)


    1 add separately

    10× PCR Buffer 2 10 20 40 60 80

    10× Mg++ (25mM) stock 1.2 6 12 24 36 48

    10× dNTPs (2mM) stock 2 10 20 40 60 80

    10× E1 primer stock (2.5 2 10 20 40 60 80


    pmoles/μl) stock
    10× E2 primer stock (2.5 2 10 20 40 60 80
    pmoles/μl) stock
    Taq DNA polymerase
    0.1 0.5 1 2 3 4
    Double-distilled H2O 9.7 48.5 97 194 291 388

    total volume 20

    Reaction mixes for MAT1 amplifications (50-μl


    reaction volumes)

    1x 5 10 20 30 40

    Target DNA (1-20 ng/μl) 2 add separately

    10× PCR Buffer 5 25 50 100 150 200

    10× Mg++ (25mM) stock 5 25 50 100 150 200

    10× dNTPs (2mM) stock 5 25 50 100 150 200

    10× GFmat1a primer stock 5 25 50 100 150 200


    (1uM/) stock
    10× GFmat1b primer stock 5 25 50 100 150 200
    (1uM/) stock
    Taq DNA polymerase
    0.25 1.25 2.5 5 7.5 10

    Double-distilled H2O
    22.75 113.75 227.5 455 682.5 910

    total volume 50

    Reaction mixes for MAT2 amplifications (50-μl


    reaction volumes)

    1x 5 10 20 30 40

    Target DNA (1-20 ng/μl) 2 add separately

    10× PCR Buffer 5 25 50 100 150 200

    10× Mg++ (25mM) stock 5 25 50 100 150 200

    10× dNTPs (2mM) stock


    5 25 50 100 150 200
    10× GFmat2c primer stock 5 25 50 100 150 200
    (1uM/) stock
    10× GFmat2d primer stock
    5 25 50 100 150 200
    (1uM/) stock
    Taq DNA polymerase
    0.25 1.25 2.5 5 7.5 10

    Double-distilled H2O 22.75 113.75 227.5 455 682.5 910

    total volume 50
    50 60 70 80 90 100

    100 120 140 160 180 200

    60 72 84 96 108 120

    100 120 140 160 180 200

    100 120 140 160 180 200

    100 120 140 160 180 200

    5 6 7 8 9 10

    485 582 679 776 873 970

    50 60 70 80 90 100

    100 120 140 160 180 200

    60 72 84 96 108 120

    100 120 140 160 180 200

    100 120 140 160 180 200

    100 120 140 160 180 200

    5 6 7 8 9 10
    485 582 679 776 873 970

    50 60 70 80 90 100

    250 300 350 400 450 500

    250 300 350 400 450 500

    250 300 350 400 450 500

    250 300 350 400 450 500

    250 300 350 400 450 500

    12.5 15 17.5 20 22.5 25

    1137.5 1365 1592.5 1820 2047.5 2275

    50 60 70 80 90 100

    250 300 350 400 450 500

    250 300 350 400 450 500

    250 300 350 400 450 500


    250 300 350 400 450 500

    250 300 350 400 450 500

    12.5 15 17.5 20 22.5 25

    1137.5 1365 1592.5 1820 2047.5 2275

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