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Basic Conversion of Nucleic Acids and Nucleotides

    1. Conversion of nucleic acids

    (1) Moles and mass

    1 ug 1,000bp DNA = 1.52 pmol
    1 ug SV40 DNA (5,243bp) = 0.29 pmol
    1 ug FX174 DNA (5,386bp) = 0.28 pmol
    1 ug lambda phage DNA (48,502bp) = 0.03 pmol
    1 ug M13mp18/19 DNA (7,249bp) = 0.21 pmol
    1 ug pBR322 DNA (4,361bp) = 0.35 pmol
    1 ug pUC18/19 DNA (2,686bp) = 0.57 pmol
    1 pmol 1,000bp DNA = 0.66 ug
    1 pmol M13mp18/19 DNA (7,249bp) = 4.78 ug
    1 pmol SV40 DNA (5,243bp) = 3.46 ug
    1 pmol pUC18/19 DNA (2,686bp) = 1.77 ug
    1 pmol pBR322 DNA (4,361bp) = 2.88 ug
    1 pmol lambda phage DNA (48,502bp) = 32.01 ug
    1 pmol FX174 DNA (5,386bp) = 3.55 ug

    (2) Absorbance and concentration

    1 OD260 dsDNA = 50 mg/ml
    1 OD260 ssDNA = 33 mg/ml
    1 OD260 ssRNA = 40 mg/ml

    (3) Molecular weight (MW)

    Average MW of dsDNA base pair = 600 Daltons
    Average MW of ssDNA base pair = 333 Daltons
    Average molecular weight of RNA base is 340 Daltons

    (4) Concentration of nucleic acid end

    Linear DNA:pmol ends = pmol DNA´ (number of cuts ´2 + 2)
    Circular DNA:pmol ends = pmol DNA´ number of cuts ´2
    1 ug linear 1,000bp DNA = 3.04 pmol ends
    1 ug linear FX174 DNA (5,386bp) = 0.56 pmol ends
    1 ug linear pUC18/19 DNA (2,686bp) = 1.14 pmol ends
    1 ug linear SV40 DNA (5,243bp) = 0.58 pmol ends
    1 ug linear pBR322 DNA (4,361bp) = 0.7 pmol ends
    1 ug linear lambda phage DNA (48,502bp) = 0.06 pmol ends
    1 ug linear M13mp18/19 DNA (7,249bp) = 0.42 pmol ends

    2. Conversion of nucleotides

    (1) Molecular weight (MW)

    MW (Da) = 333’N (number of bases)

    (2) Concentration

    C (mmol/L or pmol/ml) = OD260 / (0.01´N) C (ng/ml) = OD260’ MW / (0.01´N)
    Tips: OD260—absorbance at 260 nm

    (3). Double-stranded DNA with a melting point of oligonucleotides:

    Shorter than 20bp double-stranded oligonucleotide: Tm = 2 (A + T) + 4 (G + C)
    Longer than 20bp double-stranded oligonucleotide: Tm = 81.5 + 16.6 ( lg[J+] ) + 0.41 (%GC) –(600/N)
    Tips: N—Length of the primer (calculate the number of bases);J+ —— Monovalent cation concentration

    (4). Protein/DNA Conversions

    1 kb DNA = 333 amino acid ≈ 3.7´104 Da
    10,000 Da Protein ≈ 270 bp DNA
    30,000 Da Protein ≈ 810 bp DNA
    50,000 Da Protein ≈ 1350 bp DNA
    100,000 Da Protein ≈ 2700 bp DNA