文献类型：专利

英文题名：Extracting nucleic acid of rape bee pollen, comprises e.g. preparing solvent by mixing sodium dodecyl sulfate extracting solution and mixture solution of chloroform/isoamylol in equal volume, carrying out wall breaking treatment on pollen

作者：GAO L;HU Y;WANG L;LI Y;LI R

机构：[1]UNIV EAST CHINA SCI & TECHNOLOGY;[2]QINGHAI RUIYUAN DRUG RES INST CO LTD;[3]INST NORTHWEST HIGHLAND SPECIES CHINESE

申请号：CN102796731-B

公开日：2014-03-05

语种：英文

收录：DERWENT

摘要：NOVELTY - Extracting nucleic acid of rape bee pollen comprises: e.g. preparing a solvent by mixing sodium dodecyl sulfate (SDS) extracting solution and mixture solution of chloroform/isoamylol in equal volume; carrying out wall breaking treatment on the rape bee pollen; homogenizing the pollen solution to obtain pollen homogeneous solution instantly; cooling the rape bee pollen cell lysis solution; transferring the supernatant to other centrifugal tube; and adding 2/3 volume of pre-cooled isopropyl alcohol obtained by a normal method in water phase. USE - The method is useful for extracting nucleic acid of rape bee pollen (claimed). ADVANTAGE - The method allows simple, rapid and economical extraction of nucleic acid without harmful residues. DETAILED DESCRIPTION - Extracting nucleic acid of rape bee pollen comprises: preparing a solvent by mixing sodium dodecyl sulfate (SDS) extracting solution and mixture solution of chloroform/isoamylol in equal volume, where the volume ratio of the chloroform to the isoamylol in the mixture solution of chloroform/isoamylol is 24:1; carrying out wall breaking treatment on the rape bee pollen to mix the rape bee pollen and the solvent in a mass volume ratio of 1:3-1:5 and then soaking for 12-24 hours to obtain the pollen solution, homogenizing the pollen solution using a homogenizer at a speed of 20000-24000 revolutions per minute (rpm) for 2-3 times with a time of 5-10 minutes to obtain pollen homogeneous solution instantly; transferring the pollen homogeneous solution to a centrifuge tube, adding the SDS extracting solution for 5-10 times to the volume of the pollen homogeneous solution and then keeping in ice bath at 60-65 degrees C for 1-1.5 hours to obtain rape bee pollen cell lysis solution; cooling the rape bee pollen cell lysis solution at room temperature, adding obtained solvent for extracting for 8-10 minutes and then centrifuging for 8-10 minutes with a speed of 10000 rpm at 0-4 degrees C to obtain supernatant; transferring the supernatant to the other centrifugal tube, and repeating the cooling step until no protein layer is existed between water and organic phases; and adding 2/3 volume of pre-cooled isopropyl alcohol obtained by a normal method in water phase and then uniformly swinging and putting obtained product for 2-3 hours at -20 degrees C or overnight to obtain liquid phase and sediment, removing the liquid phase, washing the sediment for 2-3 times by using ethanol with mass concentration of 75%, removing ethanol and naturally drying the product to obtain the bee pollen nucleic acid, weighing the dry product and then dissolving the rape bee pollen nucleic acid using double distilled water and preserving at 0-4 degrees C, and storing for a long time at 20 degrees C, where the mass volume ratio of the rape bee pollen nucleic acid to the double distilled water is 1:50-1:120.