Method for extracting and purifying semen DNA
A semen and solution technology, applied in the field of DNA extraction and purification, can solve the problems of not meeting the requirements of DNA extraction and purification, insufficient DNA purity, etc.
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Embodiment 1
[0067] Example 1. Extraction and purification of DNA in a trace amount of blood and verification of its effect
[0068] 1. Extraction and purification of DNA in trace amounts of blood and STR multiplex amplification analysis
[0069] 1. Reagents
[0070] The reagents are as follows:
[0071] Pretreatment lysate: pH is 7.4, and solvent is water, and solute is the material of following final concentration: 10mM Tris-HCl, the sodium dodecylsulfonate (SDS) of 0.5g / 100ml, the NaCl aqueous solution of the EDTA of 1mM and 50mM, 0.5 mg / ml proteinase K.
[0072]Lysis binding solution: pH is 6.4, solvent is water, and solute is the material of following final concentration: the Tris-HCl of 50mM, the guanidine isothiocyanate of 3M, the Triton X-100 of 1% (volume percentage), 0.5g / 100ml 3-(3-cholaminopropyl)-dimethylamino-1-propanesulfonic acid (CHAPS), 10 mM EDTA and 15% (volume percent) ethanol.
[0073] Washing solution I: the solvent is water, and the solute is the following final...
Embodiment 2
[0113] Example 2, Extraction and Purification of DNA in Purified Blood Spots and Validation of Effects
[0114] A 3×3mm blood spot on the FTA card was used as a biological sample, and the DNA in the sample was extracted and purified.
[0115] 1. Extraction and purification of DNA from blood spots
[0116] 1. Reagents
[0117] The reagents are as follows:
[0118] Pretreatment lysate: pH is 8.0, the solvent is water, and the solute is the substance of the following final concentration: 50mM Tris-HCl, 3g / 100ml sodium dodecylsulfonate (SDS), 50mM CDTA and 100mM NaCl aqueous solution, 0.2 mg / ml proteinase K.
[0119] Lysis binding solution: pH is 7.0, solvent is water, and solute is the material of following final concentration: the Tris-HCl of 100mM, the guanidine isothiocyanate of 5M, the Triton X-100 of 5% (volume percent), 2g / 100ml 3-(3-cholaminopropyl)-dimethylamino-1-propanesulfonic acid (CHAPS), 50 mM EDTA and 20% (volume percent) ethanol.
[0120] Washing solution I: ...
Embodiment 3
[0140] Example 3, Extraction and Purification of DNA in Purified Blood Spots and Validation of Effects
[0141] A sample in which 2 μl of anticoagulated blood was dropped on a cotton swab was used as a biological sample, and DNA in the sample was extracted and purified.
[0142] 1. Extraction and purification of DNA
[0143] 1. Reagents
[0144] The reagents are as follows:
[0145] Pretreatment lysate: pH is 8.5, and solvent is water, and solute is the material of following final concentration: 100mM phosphate (100mM NaH 2 PO 4 , 20mM NaCl, pH 8.2), 5.0g / 100ml of sodium lauryl sarcosine (SLS), 100mM of EGTA and 150mM of NaCl in water, 5.0mg / ml of proteinase K.
[0146] Lysis binding liquid: pH is 7.4, and solvent is water, and solute is the material of following final concentration: the Tris-HCl of 150mM, the guanidine thiocyanate of 8M, the Tween 20 of 10% (volume percentage), the 3- (3-cholaminopropyl)-dimethylamino-1-propanesulfonic acid (CHAPS), 100 mM EGTA and 30% (...
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