160 results about "Guanidine isothiocyanate" patented technology

The invention relates to the field of molecular biology and discloses a kit and a method for extracting DNA from histiocytes. The kit comprises a binding solution, a regulating and binding solution and a deproteinization solution, wherein the binding solution comprises 3-5M of guanidinium isothiocyanate, 5-50mM of Tris-HCl with a pH value of 3.5-5.5, 5-25 percent by weight of Ttiton-X 100 and 5-20mM of Urea; the regulating and binding solution is isopropyl alcohol; and the deproteinization solution comprises 2-6M of guanidine hydrochloride, 5-50mM of Tris-HCl with a pH value of 6.5-8.0 and 5-50 percent by weight of absolute ethyl alcohol. The kit also comprises a pretreatment buffer solution, a splitting solution, a magnetic bead, a rinsing solution and an eluting solution. The DNA extracted by the kit and the method has the advantages of high content and high extraction speed, and the invention does not adopt toxic reagents and is suitable for extracting the DNA of various histiocytes.

The invention discloses a method for extracting microbial total RNA in forest soil and litter, which comprises the following steps: 1, carrying out freeze-drying on a forest soil or litter sample, grinding, and uniformly mixing; 2, adding pyrocarbonic acid diethyl ester treating water into the powdered sample, and standing over night at -70 DEG C; 3, adding glass beads, hexadecyl trimethyl ammonium bromide buffer, lauryl sodium sulfate, diatomite and phenol / chloroform / isoamyl alcohol, and uniformly mixing; 4, severely shaking the mixed liquid, and centrifuging to obtain the supernate; 5, adding guanidinium isothiocyanate into the supernate, and centrifuging to obtain the supernate; 6, adding chloroform / isoamyl alcohol into the supernate, and centrifuging to obtain the supernate; 7, addingpolyethylene glycol 6000 into the supernate, standing, centrifuging, and removing the supernate; 8, washing, dissolving, and carrying out DNA enzyme digestion to obtain an RNA solution; and 9, detecting the integrality of the RNA through agarose gel electrophoresis, and using a nucleic acid protein determinator to detect the concentration and purity of the RNA. The RNA extracted by using the method has high yield, good integrality and better purity.

The invention provides a nucleic acid releasing agent, and belongs to the technical field of nucleic acid extraction. The nucleic acid releasing agent is a liquid preparation. In the nucleic acid releasing agent, volume percentage content of a nonionic surfactant is 0.2-2%, mass volume percentage content of trehalose is 0.01-2%, mass volume concentration of protease K is 0.1-5 mg / ml, concentrationof potassium chloride is 50-200 mmol / L, the concentration of guanidinium isothiocyanate is 20-100 mmol / L, the concentration of bovine serum albumin is 0.5-10 mmol / L, the concentration of ethylene diamine tetraacetic acid is 1-10 mmol / L, and the concentration of Tris-HCl is 0.2-5 mmol / L. The nucleic acid releasing agent further comprises the balance of water. The nucleic acid releasing agent is capable of eliminating a recovering step of nucleic acid, and avoiding nucleic acid losing or polluting. The nucleic acid releasing agent can be applied for realizing on-site releasing and detecting ofthe nucleic acid.