[原创]RNA Extraction Reagent (TRIZOL)
<p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt; tab-stops: 90.25pt center 207.65pt;"><b style="mso-bidi-font-weight: normal;"><span lang="EN-US" style="FONT-SIZE: 16pt;"><font face="Times New Roman">RNA Extraction Reagent<span style="mso-spacerun: yes;"> </span>(TRIZOL)<p></p></font></span></b></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt; tab-stops: 90.25pt center 207.65pt;"><font size="3"><span lang="EN-US"><font face="Times New Roman">Catalogue</font></span><span style="FONT-FAMILY: 宋体; mso-ascii-font-family: "Times New Roman"; mso-hansi-font-family: "Times New Roman";">：</span><span lang="EN-US"><font face="Times New Roman">MN012<span style="mso-spacerun: yes;"> </span>Storage: 4</font></span><span style="FONT-FAMILY: 宋体; mso-ascii-font-family: "Times New Roman"; mso-hansi-font-family: "Times New Roman";">－</span><span lang="EN-US"><font face="Times New Roman">8 ºC</font></span></font></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt; LINE-HEIGHT: 150%;"><b style="mso-bidi-font-weight: normal;"><i style="mso-bidi-font-style: normal;"><span lang="EN-US" style="FONT-SIZE: 9pt; LINE-HEIGHT: 150%; FONT-FAMILY: Verdana;">Description:</span></i></b><span lang="EN-US" style="FONT-SIZE: 9pt; LINE-HEIGHT: 150%; FONT-FAMILY: Verdana;"><p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt; LINE-HEIGHT: 150%;"><span lang="EN-US" style="FONT-SIZE: 9pt; LINE-HEIGHT: 150%; FONT-FAMILY: Verdana;">TRIZOL is a mono-phasic reagent for the isolation of total RNA from cells and tissues. The reagent, containing phenol and guanidine isothiocyanate, extracts total RNA during lysing the cells in a single-step procedure.<p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt; LINE-HEIGHT: 150%;"><b style="mso-bidi-font-weight: normal;"><i style="mso-bidi-font-style: normal;"><span lang="EN-US" style="FONT-SIZE: 9pt; LINE-HEIGHT: 150%; FONT-FAMILY: Verdana;">Procedure:<p></p></span></i></b></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt; LINE-HEIGHT: 150%;"><i style="mso-bidi-font-style: normal;"><u><span lang="EN-US" style="FONT-SIZE: 9pt; LINE-HEIGHT: 150%; FONT-FAMILY: Verdana;">I. RNA Preparation for Cells Grown in Monolayer<p></p></span></u></i></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 21.6pt; TEXT-INDENT: -18pt; LINE-HEIGHT: 150%; mso-list: l3 level1 lfo2; tab-stops: list 21.6pt;"><span lang="EN-US" style="FONT-SIZE: 9pt; LINE-HEIGHT: 150%; FONT-FAMILY: Verdana; mso-fareast-font-family: Verdana; mso-bidi-font-family: Verdana;"><span style="mso-list: Ignore;">1.<span style="FONT: 7pt "Times New Roman";"> </span></span></span><span lang="EN-US" style="FONT-SIZE: 9pt; LINE-HEIGHT: 150%; FONT-FAMILY: Verdana;">Lyse cells directly in a culture plate/dish/flask by adding 1 ml of TRIZOL Reagent to a 3.5<chmetcnv wst="on" unitname="cm" sourcevalue="10" hasspace="True" negative="True" numbertype="1" tcsc="0">-10 cm</chmetcnv> dish or similar cell culture devices, and passing the cell lysate several times through a pipette. <p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 21.6pt; TEXT-INDENT: -18pt; LINE-HEIGHT: 150%; mso-list: l3 level1 lfo2; tab-stops: list 21.6pt;"><span lang="EN-US" style="FONT-SIZE: 9pt; LINE-HEIGHT: 150%; FONT-FAMILY: Verdana; mso-fareast-font-family: Verdana; mso-bidi-font-family: Verdana;"><span style="mso-list: Ignore;">2.<span style="FONT: 7pt "Times New Roman";"> </span></span></span><span lang="EN-US" style="FONT-SIZE: 9pt; LINE-HEIGHT: 150%; FONT-FAMILY: Verdana;">Incubate the homogenized samples for 5 minutes at room temperature (RT) to permit the complete dissociation of nucleoprotein complexes. <p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 21.6pt; TEXT-INDENT: -18pt; LINE-HEIGHT: 150%; mso-list: l3 level1 lfo2; tab-stops: list 21.6pt;"><span lang="EN-US" style="FONT-SIZE: 9pt; LINE-HEIGHT: 150%; FONT-FAMILY: Verdana; mso-fareast-font-family: Verdana; mso-bidi-font-family: Verdana;"><span style="mso-list: Ignore;">3.<span style="FONT: 7pt "Times New Roman";"> </span></span></span><span lang="EN-US" style="FONT-SIZE: 9pt; LINE-HEIGHT: 150%; FONT-FAMILY: Verdana;">Add 0.2 mL of chloroform per 1 mL of TRIZOL Reagent. Shake tubes vigorously for 15 seconds and incubate them at RT for 3 minutes. (see note 1) <p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 21.6pt; TEXT-INDENT: -18pt; LINE-HEIGHT: 150%; mso-list: l3 level1 lfo2; tab-stops: list 21.6pt;"><span lang="EN-US" style="FONT-SIZE: 9pt; LINE-HEIGHT: 150%; FONT-FAMILY: Verdana; mso-fareast-font-family: Verdana; mso-bidi-font-family: Verdana;"><span style="mso-list: Ignore;">4.<span style="FONT: 7pt "Times New Roman";"> </span></span></span><span lang="EN-US" style="FONT-SIZE: 9pt; LINE-HEIGHT: 150%; FONT-FAMILY: Verdana;">Centrifuge the samples at 10,000 × g for 10 minutes at RT. <p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 21.6pt; TEXT-INDENT: -18pt; LINE-HEIGHT: 150%; mso-list: l3 level1 lfo2; tab-stops: list 21.6pt;"><span lang="EN-US" style="FONT-SIZE: 9pt; LINE-HEIGHT: 150%; FONT-FAMILY: Verdana; mso-fareast-font-family: Verdana; mso-bidi-font-family: Verdana;"><span style="mso-list: Ignore;">5.<span style="FONT: 7pt "Times New Roman";"> </span></span></span><span lang="EN-US" style="FONT-SIZE: 9pt; LINE-HEIGHT: 150%; FONT-FAMILY: Verdana;">Transfer the aqueous phase to a fresh tube. Precipitate the RNA from the aqueous phase by mixing with isopropyl alcohol. Use 0.5 mL of isopropyl alcohol per 1 ml of TRIZOL Reagent used for the initial homogenization.<p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 21.6pt; TEXT-INDENT: -18pt; LINE-HEIGHT: 150%; mso-list: l3 level1 lfo2; tab-stops: list 21.6pt;"><span lang="EN-US" style="FONT-SIZE: 9pt; LINE-HEIGHT: 150%; FONT-FAMILY: Verdana; mso-fareast-font-family: Verdana; mso-bidi-font-family: Verdana;"><span style="mso-list: Ignore;">6.<span style="FONT: 7pt "Times New Roman";"> </span></span></span><span lang="EN-US" style="FONT-SIZE: 9pt; LINE-HEIGHT: 150%; FONT-FAMILY: Verdana;">Incubate samples at RT for 10 minutes and centrifuge at no more than 10,000 × g for 10 minutes at 2 to <chmetcnv wst="on" unitname="ﾰC" sourcevalue="8" hasspace="False" negative="False" numbertype="1" tcsc="0">8°C</chmetcnv>.<p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 21.6pt; TEXT-INDENT: -18pt; LINE-HEIGHT: 150%; mso-list: l3 level1 lfo2; tab-stops: list 21.6pt;"><span lang="EN-US" style="FONT-SIZE: 9pt; LINE-HEIGHT: 150%; FONT-FAMILY: Verdana; mso-fareast-font-family: Verdana; mso-bidi-font-family: Verdana;"><span style="mso-list: Ignore;">7.<span style="FONT: 7pt "Times New Roman";"> </span></span></span><span lang="EN-US" style="FONT-SIZE: 9pt; LINE-HEIGHT: 150%; FONT-FAMILY: Verdana;">Remove the supernatant. Wash the RNA pellet once with 75% ethanol.<p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 21.6pt; TEXT-INDENT: -18pt; LINE-HEIGHT: 150%; mso-list: l3 level1 lfo2; tab-stops: list 21.6pt;"><span lang="EN-US" style="FONT-SIZE: 9pt; LINE-HEIGHT: 150%; FONT-FAMILY: Verdana; mso-fareast-font-family: Verdana; mso-bidi-font-family: Verdana;"><span style="mso-list: Ignore;">8.<span style="FONT: 7pt "Times New Roman";"> </span></span></span><span lang="EN-US" style="FONT-SIZE: 9pt; LINE-HEIGHT: 150%; FONT-FAMILY: Verdana;">Centrifuge at 10,000 × g for 5 minutes at <chmetcnv wst="on" unitname="ﾰC" sourcevalue="4" hasspace="False" negative="False" numbertype="1" tcsc="0">4°C</chmetcnv>. <p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 21.6pt; TEXT-INDENT: -18pt; LINE-HEIGHT: 150%; mso-list: l3 level1 lfo2; tab-stops: list 21.6pt;"><span lang="EN-US" style="FONT-SIZE: 9pt; LINE-HEIGHT: 150%; FONT-FAMILY: Verdana; mso-fareast-font-family: Verdana; mso-bidi-font-family: Verdana;"><span style="mso-list: Ignore;">9.<span style="FONT: 7pt "Times New Roman";"> </span></span></span><span lang="EN-US" style="FONT-SIZE: 9pt; LINE-HEIGHT: 150%; FONT-FAMILY: Verdana;">Briefly dry the RNA pellet (see note 2). Dissolve RNA in RNase-free water.<p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 3.6pt; LINE-HEIGHT: 150%;"><span lang="EN-US" style="FONT-SIZE: 9pt; LINE-HEIGHT: 150%; FONT-FAMILY: Verdana;"><p> </p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt; LINE-HEIGHT: 150%;"><i style="mso-bidi-font-style: normal;"><u><span lang="EN-US" style="FONT-SIZE: 9pt; LINE-HEIGHT: 150%; FONT-FAMILY: Verdana;">I. RNA Preparation for Cells Grown in Suspension<p></p></span></u></i></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 21.6pt; TEXT-INDENT: -18pt; LINE-HEIGHT: 150%; mso-list: l1 level1 lfo3; tab-stops: list 21.6pt;"><span lang="EN-US" style="FONT-SIZE: 9pt; LINE-HEIGHT: 150%; FONT-FAMILY: Verdana; mso-fareast-font-family: Verdana; mso-bidi-font-family: Verdana;"><span style="mso-list: Ignore;">1.<span style="FONT: 7pt "Times New Roman";"> </span></span></span><span lang="EN-US" style="FONT-SIZE: 9pt; LINE-HEIGHT: 150%; FONT-FAMILY: Verdana;">Pellet cells by centrifugation. <p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 21.6pt; TEXT-INDENT: -18pt; LINE-HEIGHT: 150%; mso-list: l1 level1 lfo3; tab-stops: list 21.6pt;"><span lang="EN-US" style="FONT-SIZE: 9pt; LINE-HEIGHT: 150%; FONT-FAMILY: Verdana; mso-fareast-font-family: Verdana; mso-bidi-font-family: Verdana;"><span style="mso-list: Ignore;">2.<span style="FONT: 7pt "Times New Roman";"> </span></span></span><span lang="EN-US" style="FONT-SIZE: 9pt; LINE-HEIGHT: 150%; FONT-FAMILY: Verdana;">Lyse cells in TRIZOL Reagent by repetitive pipetting.<p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 21.6pt; TEXT-INDENT: -18pt; LINE-HEIGHT: 150%; mso-list: l1 level1 lfo3; tab-stops: list 21.6pt;"><span lang="EN-US" style="FONT-SIZE: 9pt; LINE-HEIGHT: 150%; FONT-FAMILY: Verdana; mso-fareast-font-family: Verdana; mso-bidi-font-family: Verdana;"><span style="mso-list: Ignore;">3.<span style="FONT: 7pt "Times New Roman";"> </span></span></span><span lang="EN-US" style="FONT-SIZE: 9pt; LINE-HEIGHT: 150%; FONT-FAMILY: Verdana;">Continue I) step 2-9.<p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 3.6pt; LINE-HEIGHT: 150%;"><span lang="EN-US" style="FONT-SIZE: 9pt; LINE-HEIGHT: 150%; FONT-FAMILY: Verdana;"><p> </p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt; LINE-HEIGHT: 150%;"><i style="mso-bidi-font-style: normal;"><u><span lang="EN-US" style="FONT-SIZE: 9pt; LINE-HEIGHT: 150%; FONT-FAMILY: Verdana;">III. RNA Preparation for Tissues<p></p></span></u></i></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 21.6pt; TEXT-INDENT: -18pt; LINE-HEIGHT: 150%; mso-list: l0 level1 lfo4; tab-stops: list 21.6pt;"><span lang="EN-US" style="FONT-SIZE: 9pt; LINE-HEIGHT: 150%; FONT-FAMILY: Verdana; mso-fareast-font-family: Verdana; mso-bidi-font-family: Verdana;"><span style="mso-list: Ignore;">1.<span style="FONT: 7pt "Times New Roman";"> </span></span></span><span lang="EN-US" style="FONT-SIZE: 9pt; LINE-HEIGHT: 150%; FONT-FAMILY: Verdana;">Homogenize tissue samples in 1 mL of TRIZOL Reagent per 50-100 mg of tissue using a glass-Teflon or power homogenizer. <p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 21.6pt; TEXT-INDENT: -18pt; LINE-HEIGHT: 150%; mso-list: l0 level1 lfo4; tab-stops: list 21.6pt;"><span lang="EN-US" style="FONT-SIZE: 9pt; LINE-HEIGHT: 150%; FONT-FAMILY: Verdana; mso-fareast-font-family: Verdana; mso-bidi-font-family: Verdana;"><span style="mso-list: Ignore;">2.<span style="FONT: 7pt "Times New Roman";"> </span></span></span><span lang="EN-US" style="FONT-SIZE: 9pt; LINE-HEIGHT: 150%; FONT-FAMILY: Verdana;">Continue I) step 2-9<p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt; LINE-HEIGHT: 150%;"><b style="mso-bidi-font-weight: normal;"><i style="mso-bidi-font-style: normal;"><span lang="EN-US" style="FONT-SIZE: 9pt; LINE-HEIGHT: 150%; FONT-FAMILY: Verdana;">Notes:<p></p></span></i></b></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 12.6pt; TEXT-INDENT: -12.6pt; LINE-HEIGHT: 150%; mso-list: l2 level1 lfo1; tab-stops: list 12.6pt;"><span lang="EN-US" style="FONT-SIZE: 9pt; LINE-HEIGHT: 150%; FONT-FAMILY: Verdana; mso-fareast-font-family: Verdana; mso-bidi-font-family: Verdana;"><span style="mso-list: Ignore;">1.<span style="FONT: 7pt "Times New Roman";"> </span></span></span><span lang="EN-US" style="FONT-SIZE: 9pt; LINE-HEIGHT: 150%; FONT-FAMILY: Verdana;">DO NOT vortex the samples, which will cause DNA contamination.<p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt; tab-stops: 90.25pt center 207.65pt;"><span lang="EN-US" style="FONT-SIZE: 9pt; FONT-FAMILY: Verdana;">DO NOT over-dry the RNA, which will decrease its solubility.</span><b style="mso-bidi-font-weight: normal;"><span lang="EN-US" style="FONT-SIZE: 16pt;"><p></p></span></b></p> <p><span style="FONT-SIZE: 12pt; FONT-FAMILY: 宋体;">迈晨科技（北京）有限公司竭诚为您服务 ！<span lang="EN-US"><p></p></span></span></p><p><span style="FONT-SIZE: 12pt; FONT-FAMILY: 宋体;">详情请登录网站：<span lang="EN-US">www.macgene.com</span>，联系电话：<span lang="EN-US">010-82057786<p></p></span></span></p> 迈晨科技（北京）有限公司竭诚为您服务 ！详情请登录网站：www.macgene.com，联系电话：010-82057786
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