利用trizol_試劑和液氮提取大鼠胰腺高質(zhì)量總rna

1、利用TRIzol_試劑和液氮提取大鼠胰腺高質(zhì)量總RNA.txt“我羨慕內(nèi)些老人羨慕他們手牽手一直走到最后。━交話費(fèi)的時(shí)候，才發(fā)現(xiàn)自己的話那么值錢。第30卷第5期西安交通大學(xué)學(xué)報(bào)(醫(yī)學(xué)版)Vol.30No.52009年10月JournalofXipanJiaotongUniversity(MedicalSciences)Oct.2009利用TRIzol試劑和液氮提取大鼠胰腺高質(zhì)量總RNA李冬民12任吳超12王璇12王飛苗12高玉12韓燕1

2、2寧啟蘭12宋天保13呂社民12(西安交通大學(xué)醫(yī)學(xué)院:1.環(huán)境與疾病相關(guān)基因教育部重點(diǎn)實(shí)驗(yàn)室2.遺傳學(xué)與分子生物學(xué)系3.人體解剖學(xué)與組織胚胎學(xué)系陜西西安710061)摘要:目的建立胰腺高質(zhì)量總RNA快速、經(jīng)濟(jì)、穩(wěn)定的提取方法。方法應(yīng)用TRIzol試劑和液氮提取大鼠胰腺總RNA通過總RNA含量和A260280比值的測定及10gL非變性瓊脂糖凝膠電泳評估總RNA的質(zhì)量并用RT2PCR檢2.DepartmentofGeicsMolecular

3、BiologyMedicalSchoolofXipanJiaotongUniversityXipan7100613.DepartmentofHumanAnatomyHistologyEmbryologyMedicalSchoolofXipanJiaotongUniversityXipan710061China)ABSTRACT:ObjectiveToestablishaquickeconomicalreproduciblemethodf

4、high2qualityRNAextractionfrompancreas.MethodsWeutilizedTRIzolReagentliquidnitrogentoisolatetotalRNAfromtheratpancreas.TheRNAqualitywasdeterminedbydetectionofitscontentopticdensity(A)at260280nmelectrophesisin1%non2denatur

5、edagarosegel.Thenreversetranion2polymerasechainreaction(RT2PCR)wasperfmedtodetectexpressionofthepancreas2specificgenes.ResultsThecontentofthetotalRNAextractedfromtheratpancreasreached36μgmgpancreatictissuesA260280ratiowa

6、s1.751.89.ElectrophesisofthetotalRNAshowed28S18SrRNAbswithclearsmearbetweenthem.TheRT2PCRproductsofpancreas2specificgenesincludinginsulin1glucagonα2amylasehousekeepinggeneβ2actinallexhibitedclearbson1%agarosegelwhichwere

7、locatedintheexpectedpositionsrespectively.ConclusionTheseresultssuggestthatwehavesuccessfullyisolatedthehigh2qualityintactRNAfromtheratpancreaswithTRIzolReagentliquidnitrogen.TheextractedtotalRNAcanbeusedinRT2PCRfpancrea