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A kind of buffer solution for coating nucleic acid primer and preparation method of coating nucleic acid primer

A technology of buffer solution and buffer solution, which is applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of high material consumption cost, complex process technology, high reagent cost, etc., to reduce process complexity and process steps, cheap effect

Active Publication Date: 2020-03-17
ZHENGZHOU KODIA BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the existing process technology takes too long, with an average time-consuming of about 4 to 5 days, and the process technology is too complicated. Repeated plate washing steps, the average process requires each plate to be washed about 15 times, and the existing The cost of process technology material consumption is too high, and the cost of reagents used for coating is too high, which needs to be imported and expensive

Method used

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  • A kind of buffer solution for coating nucleic acid primer and preparation method of coating nucleic acid primer
  • A kind of buffer solution for coating nucleic acid primer and preparation method of coating nucleic acid primer
  • A kind of buffer solution for coating nucleic acid primer and preparation method of coating nucleic acid primer

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preparation example Construction

[0039] The present invention also provides a method for preparing a coated nucleic acid primer, comprising the following steps:

[0040] A) dissolving the nucleic acid primer in the TE buffer solution to obtain a mixed solution;

[0041] After mixing disodium hydrogen phosphate, sodium dihydrogen phosphate, polylysine, sodium chloride, preservatives and water, a buffer solution is obtained;

[0042] B) After mixing the mixed solution obtained in the above steps and the buffer solution again, the coating mixed solution is obtained;

[0043] C) Put the coating mixture obtained in the above steps into a coating plate and perform coating to obtain coated nucleic acid primers.

[0044] In the preparation method of the present invention, the specific preferred scheme of the raw material, and the preferred numerical value of the number of parts added are consistent with the specific preferred scheme of the components in the aforementioned buffer solution, and the preferred numerical...

Embodiment 1

[0055] Application of coating buffer in the field of plate chemiluminescence;

[0056] Step 1: First design a nucleic acid sequence of a coating primer (5'-NH2-GAGCGGATATATGCTTAGGAAT-3'), synthesize the sequence and dissolve it in a TE buffer solution with a pH of 8.0, and dilute the primer to 3ug / mL;

[0057] Step 2: prepare coating buffer, according to the following concentration ratio, disodium hydrogen phosphate 5.6g / L, sodium dihydrogen phosphate 2.8g / L, sodium chloride 0.6g / L, sodium sulfate 17g / L , 28g / L poly-lysine, 0.06g / L preservative and deionized water, pay attention, add sodium phosphate buffer first, then add poly-lysine, and add easily soluble sodium sulfate after the solution is clarified and sodium chloride reagent, and magnetically stir at room temperature for 2 hours in the dark;

[0058] Step 3: Mix the primers dissolved in TE buffer and the prepared coating buffer in equal proportions to ensure that the final concentration of primers is 1.5 μg / mL;

[005...

Embodiment 2

[0083] Magnetic bead coating process:

[0084] 1. Add 500mg of amino (PVA) magnetic beads to a 1.5ml EP tube, wash 3 times with 1X pBS; add 0.1M MES for 3min;

[0085] 2. Adsorb the magnetic beads, pour off the MES solution, and add 1ml of the coating buffer solution provided by the present invention (according to the following concentration ratio, 5.5g / L disodium hydrogen phosphate, 3.0g / L sodium dihydrogen phosphate, 0.7 g / L sodium chloride, 18g / L sodium sulfate, 30g / L polylysine, 0.09g / L preservative and deionized water), then add primer 100pmol, and react for 2 to 3 hours;

[0086] 3. Add magnetic bead blocking agent (JSR, Cat#65293), end the reaction, adsorb the magnetic beads, and pour off the blocking agent;

[0087] 4. Wash three times with 1XPBS, dissolve the magnetic beads in 1X TE buffer, and set aside at 4°C.

[0088] So far, the coating process of the capture magnetic beads is completed.

[0089] Design a capture probe sequence as follows, see Table 5:

[0090...

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Abstract

The invention provides a buffer solution used for coating a nucleic acid primer. The buffer solution comprises the following components in mass-volume concentration: 5.3-5.6g / L of disodium hydrogen phosphate, 2.5-3.0g / L of sodium dihydrogen phosphate, 0.5-0.8g / L of sodium chloride, 25-32g / L of polylysine, 0.03-0.09g / L of a preservative and water. The buffer solution for coating is adopted, so that the integral time for coating the nucleic acid primer can be effectively reduced, complex process steps can be reduced, and cost benefits are greatly reduced through a one-step process. The buffer solution for coating greatly reduces the raw material cost, so that the price of the final product is lower.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a buffer for coating nucleic acid primers and a preparation method for coating nucleic acid primers. Background technique [0002] With the rapid development of molecular biology and the extensive application of biophysical technology, nucleic acid hybridization is used in the diagnosis of tumors or cancers, infections such as viruses and bacteria, and the detection of various genetic diseases. In the prior art, nucleic acid primers are coated on microwell plates, such as polystyrene plastic plates, for the diagnosis of tumors or cancer, infections such as viruses and bacteria, and the detection of various genetic diseases. [0003] Nucleic acid primer, also known as primer, is a small piece of single-stranded DNA or RNA, which acts as the starting point of DNA replication and as the starting point for the extension of each polynucleotide chain during the nucleic acid synthesis react...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6806
CPCC12Q1/6806C12Q2565/537C12Q2563/143C12Q2563/149
Inventor 张鹭鹭李先坤张爱国
Owner ZHENGZHOU KODIA BIOTECHNOLOGY CO LTD
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