echemi logo
Product
  • Product
  • Supplier
  • Inquiry
    Home > Biochemistry News > Biotechnology News > The phosphorus method determines the content of nucleic acids.

    The phosphorus method determines the content of nucleic acids.

    • Last Update: 2020-10-21
    • Source: Internet
    • Author: User
    Search more information of high quality chemicals, good prices and reliable suppliers, visit www.echemi.com

    "< the principles and methods of > nucleic acid content <


    1. Experimental equipment

    thermostat water bath, 721 d'photonometer

    < "p style" text-align: left; ">2. Experiment reagent

    (1) Standard phosphorus solution: the potassium dehydrophosphate at 110 degrees C baked to constant weight, accurately called 0.8775g dissolved in a small amount of distilled water, transferred to 500 ml capacity bottle, add 5 ml. 5mol/L sulphate solution and chloroform drops, diluted with distilled water to the scale. This solution contains 400 μg of phosphorus per 1m1 and is accurately diluted 20 times (20 μg/m1) when in use.

    (2) fixed phosphorus reagent

    (1) 17% sulphuric acid: 17m1 sulphuric acid (relative density l.84) is slowly added to 83m1 water.

    (2) 2.5% ammonium diphate solution: 2.5g ammonium phthalate dissolved in 100ml water.

    (3) 10% assulphic acid solution: 10g asseptic acid dissolved in 100ml water, stored in a brown bottle, the solution is pale yellow and can be used, dark yellow or even brown is invalid.

    Mix the above three solutions with water in the following proportions: solution (1): solution (2): solution (3): water: 1:1:1:2 (V: V)

    (3) 5% ammonia, 27% sulphuric acid "

    1. The drawing of the phosphorus standard curve

    takes dryingtest tube7 numbers, as shown in the table to add reagents.

    4"

    reagents

    tube number

    0

    1

    .

    2

    <"text-left;">3

    5

    6

    0.0

    0.05

    <"text-align: left;">0.1

    0.2

    0.3

    .

    0.4

    .

    0.5

    .

    distilled water,"ml

    3.0

    <"text-right ">2.95

    0.9

    2.8

    .

    0.7

    .

    <: left;">2.6

    <:left;">2.5

    phosphorus reagents, ml

    < left;">3.0

    3.0

    .

    3.0

    .

    3.0

    <:left;">3.0

    <. > 3.0

    3.0

    A660

    .

    <"text-> < p"text-align:left;">

    ."

    < the p style is "text-aligned: left;" >

    plus bi shakes well, keeps warm 10min in a 45c water bath, cools, and measures the absorbance at 660nm with zero tube. The phosphorus content is the horizontal coordinate and the absorbance is the ordinate.

    2. The determination of total phosphorus

    called crude nucleic acid 0.1g, dissolved with a small amount of water (if insoluble, can be dripped plus 5% ammonia to pH7.0), to be fully dissolved, moved to a 50 ml capacity bottle, add water to the scale (this solution contains sample 2 mg/m1), that is, as a nucleic acid solution.


    cooling down, the digestive fluid into a 100 ml capacity bottle, with a small amount of distilled water wash test tube twice, washing liquid poured into the capacity bottle, and then add steaming water to the scale, mixed after absorbing 3m1 solution into the test tube, plus 3m1 fixed phosphorus reagent, 45c water bath insulation 10min after removal, test A660.

    3. Inorganic phosphorus determination

    absorb nucleic acid solution 1 ml, placed in a 100 ml capacity bottle, add water to scale, mix and absorb 3.0 ml test tube, add phosphorus reagent 3.0m1, 45 degrees C water bath insulation 10min after removal, test A660.

    4. Results the

    total phosphorus A660- inorganic phosphorus A660 - organophosphorus A660

    was detected from the standard curve of the number of organophosphorus micrograms (X), the nucleic acid percentage content of the pressed calculation sample

    <span s.

    This article is an English version of an article which is originally in the Chinese language on echemi.com and is provided for information purposes only. This website makes no representation or warranty of any kind, either expressed or implied, as to the accuracy, completeness ownership or reliability of the article or any translations thereof. If you have any concerns or complaints relating to the article, please send an email, providing a detailed description of the concern or complaint, to service@echemi.com. A staff member will contact you within 5 working days. Once verified, infringing content will be removed immediately.

    Contact Us

    The source of this page with content of products and services is from Internet, which doesn't represent ECHEMI's opinion. If you have any queries, please write to service@echemi.com. It will be replied within 5 days.

    Moreover, if you find any instances of plagiarism from the page, please send email to service@echemi.com with relevant evidence.