国产三级精品三级在专区_琪琪av色原伊人大芭蕉_免费A级黄毛片_精品国产AV色欲果冻传媒

Qiagen QIAprep Spin Miniprep Kit 27106

注意事(shi)項：
1．試劑盒從冷藏環境中取出應在室溫平衡(heng)15-30分鐘后方可使用，酶(mei)標包被(bei)板開(kai)封(feng)后如(ru)未(wei)用完，板條應裝入密(mi)封(feng)袋中保存。
2．濃洗滌液可(ke)能會有結(jie)晶析出，稀釋時(shi)可(ke)在水浴中加溫助溶，洗滌時(shi)不影響(xiang)結(jie)果。
3．各步加樣(yang)(yang)均應使(shi)用加樣(yang)(yang)器，并經常校對其準(zhun)確性，以(yi)避免(mian)試(shi)驗誤差。一次加樣(yang)(yang)時(shi)間最好(hao)控制在5分鐘內，如標本(ben)數量多，推薦使(shi)用排槍(qiang)加樣(yang)(yang)。
4．請每次測定(ding)的(de)同時做標準曲線，最(zui)好做復孔。如(ru)標本中待測物質含量過高（樣本OD值(zhi)大(da)于標準品孔孔的(de)OD值(zhi)），請先用樣品稀釋(shi)(shi)液稀釋(shi)(shi)一定(ding)倍(bei)數（n倍(bei)）后再測定(ding)，計(ji)算(suan)時請最(zui)后乘以總(zong)稀釋(shi)(shi)倍(bei)數（×n×5）。
5．封板膜(mo)只限(xian)一(yi)次性使用(yong)，以避(bi)免交叉污染。
6．底(di)物請避光保(bao)存。
7．嚴格按照說明書的(de)操作(zuo)進行，試驗結果判定(ding)必須以酶標儀(yi)讀(du)數為準.
8．所(suo)有樣品，洗滌液和各種廢(fei)棄(qi)物都應按傳染(ran)物處理。
9．本試劑不同批號組分不得混(hun)用。
10. 如與英文說明(ming)書有(you)異(yi)，以英文說明(ming)書為準。

Qiagen QIAprep Spin Miniprep Kit 27106

操作步(bu)驟(zou)
1.標(biao)(biao)(biao)(biao)準(zhun)(zhun)品的稀(xi)釋(shi)與加(jia)(jia)樣(yang)：在酶標(biao)(biao)(biao)(biao)包被板(ban)上(shang)設標(biao)(biao)(biao)(biao)準(zhun)(zhun)品孔(kong)(kong)(kong)10孔(kong)(kong)(kong)，在、第(di)(di)(di)(di)(di)(di)(di)(di)(di)二孔(kong)(kong)(kong)中(zhong)分(fen)別(bie)加(jia)(jia)標(biao)(biao)(biao)(biao)準(zhun)(zhun)品100μl，然(ran)后(hou)(hou)在、第(di)(di)(di)(di)(di)(di)(di)(di)(di)二孔(kong)(kong)(kong)中(zhong)加(jia)(jia)標(biao)(biao)(biao)(biao)準(zhun)(zhun)品稀(xi)釋(shi)液(ye)(ye)(ye)50μl，混勻；然(ran)后(hou)(hou)從(cong)孔(kong)(kong)(kong)、第(di)(di)(di)(di)(di)(di)(di)(di)(di)二孔(kong)(kong)(kong)中(zhong)各取(qu)100μl分(fen)別(bie)加(jia)(jia)到第(di)(di)(di)(di)(di)(di)(di)(di)(di)三(san)孔(kong)(kong)(kong)和(he)第(di)(di)(di)(di)(di)(di)(di)(di)(di)四(si)孔(kong)(kong)(kong)，再在第(di)(di)(di)(di)(di)(di)(di)(di)(di)三(san)、第(di)(di)(di)(di)(di)(di)(di)(di)(di)四(si)孔(kong)(kong)(kong)分(fen)別(bie)加(jia)(jia)標(biao)(biao)(biao)(biao)準(zhun)(zhun)品稀(xi)釋(shi)液(ye)(ye)(ye)50μl，混勻；然(ran)后(hou)(hou)在第(di)(di)(di)(di)(di)(di)(di)(di)(di)三(san)孔(kong)(kong)(kong)和(he)第(di)(di)(di)(di)(di)(di)(di)(di)(di)四(si)孔(kong)(kong)(kong)中(zhong)先各取(qu)50μl棄掉，再各取(qu)50μl分(fen)別(bie)加(jia)(jia)到第(di)(di)(di)(di)(di)(di)(di)(di)(di)五(wu)(wu)、第(di)(di)(di)(di)(di)(di)(di)(di)(di)六孔(kong)(kong)(kong)中(zhong)，再在第(di)(di)(di)(di)(di)(di)(di)(di)(di)五(wu)(wu)、第(di)(di)(di)(di)(di)(di)(di)(di)(di)六孔(kong)(kong)(kong)中(zhong)分(fen)別(bie)加(jia)(jia)標(biao)(biao)(biao)(biao)準(zhun)(zhun)品稀(xi)釋(shi)液(ye)(ye)(ye)50ul，混勻；混勻后(hou)(hou)從(cong)第(di)(di)(di)(di)(di)(di)(di)(di)(di)五(wu)(wu)、第(di)(di)(di)(di)(di)(di)(di)(di)(di)六孔(kong)(kong)(kong)中(zhong)各取(qu)50μl分(fen)別(bie)加(jia)(jia)到第(di)(di)(di)(di)(di)(di)(di)(di)(di)七(qi)、第(di)(di)(di)(di)(di)(di)(di)(di)(di)八(ba)孔(kong)(kong)(kong)中(zhong)，再在第(di)(di)(di)(di)(di)(di)(di)(di)(di)七(qi)、第(di)(di)(di)(di)(di)(di)(di)(di)(di)八(ba)孔(kong)(kong)(kong)中(zhong)分(fen)別(bie)加(jia)(jia)標(biao)(biao)(biao)(biao)準(zhun)(zhun)品稀(xi)釋(shi)液(ye)(ye)(ye)50μl，混勻后(hou)(hou)從(cong)第(di)(di)(di)(di)(di)(di)(di)(di)(di)七(qi)、第(di)(di)(di)(di)(di)(di)(di)(di)(di)八(ba)孔(kong)(kong)(kong)中(zhong)分(fen)別(bie)取(qu)50μl加(jia)(jia)到第(di)(di)(di)(di)(di)(di)(di)(di)(di)九(jiu)、第(di)(di)(di)(di)(di)(di)(di)(di)(di)十(shi)孔(kong)(kong)(kong)中(zhong)，再在第(di)(di)(di)(di)(di)(di)(di)(di)(di)九(jiu)第(di)(di)(di)(di)(di)(di)(di)(di)(di)十(shi)孔(kong)(kong)(kong)分(fen)別(bie)加(jia)(jia)標(biao)(biao)(biao)(biao)準(zhun)(zhun)品稀(xi)釋(shi)液(ye)(ye)(ye)50μl，混勻后(hou)(hou)從(cong)第(di)(di)(di)(di)(di)(di)(di)(di)(di)九(jiu)第(di)(di)(di)(di)(di)(di)(di)(di)(di)十(shi)孔(kong)(kong)(kong)中(zhong)各取(qu)50μl棄掉。（稀(xi)釋(shi)后(hou)(hou)各孔(kong)(kong)(kong)加(jia)(jia)樣(yang)量(liang)都為50μl，濃度分(fen)別(bie)為6 pmol/L，4 pmol/L ，2 pmol/L，1 pmol/L，0.5 pmol/L）。
2.加(jia)(jia)樣(yang)(yang)：分別設空白(bai)孔(kong)（空白(bai)對照孔(kong)不加(jia)(jia)樣(yang)(yang)品及(ji)酶標試劑(ji)，其(qi)余各步(bu)操作相(xiang)同）、待(dai)(dai)測(ce)樣(yang)(yang)品孔(kong)。在酶標包(bao)被板(ban)上待(dai)(dai)測(ce)樣(yang)(yang)品孔(kong)中先加(jia)(jia)樣(yang)(yang)品稀(xi)(xi)釋液40μl，然后再(zai)加(jia)(jia)待(dai)(dai)測(ce)樣(yang)(yang)品10μl（樣(yang)(yang)品最終稀(xi)(xi)釋度為5倍(bei)）。加(jia)(jia)樣(yang)(yang)將樣(yang)(yang)品加(jia)(jia)于(yu)酶標板(ban)孔(kong)底部(bu)，盡量不觸及(ji)孔(kong)壁，輕輕晃動混(hun)勻。
3.溫(wen)育：用封板(ban)膜封板(ban)后置37℃溫(wen)育30分鐘(zhong)。
4.配(pei)液：將30（48T的20倍）倍濃縮洗滌液用(yong)蒸餾水30（48T的20倍）倍稀釋后備用(yong)。
5.洗滌：小心揭(jie)掉封板膜，棄(qi)去液(ye)體，甩干(gan)，每孔加滿(man)洗滌液(ye)，靜置30秒后棄(qi)去，如此重復5次，拍干(gan)。
6.加酶：每孔(kong)加入(ru)酶標試劑50μl，空白孔(kong)除外。
7.溫(wen)育：操作同3。
8.洗滌(di)：操作(zuo)同5。
9.顯色(se)(se)：每孔先加入(ru)顯色(se)(se)劑A50μl，再加入(ru)顯色(se)(se)劑B50μl，輕(qing)輕(qing)震蕩混勻，37℃避光顯色(se)(se)15分鐘.
10.終止(zhi)(zhi)：每孔加終止(zhi)(zhi)液50μl，終止(zhi)(zhi)反應（此時藍色(se)(se)立轉黃色(se)(se)）。
11.測定：以(yi)空白空調零，450nm波長依序(xu)測量各孔的吸光度（OD值）。 測定應在加終止液后15分鐘以(yi)內進行(xing)。

樣本處理及要(yao)求：
1. 血(xue)清(qing)：室(shi)溫血(xue)液自然凝固10-20分鐘，離心(xin)20分鐘左右（2000-3000轉/分）。仔細(xi)收(shou)集上清(qing)，保存過程中(zhong)如出現沉淀，應再次離心(xin)。
2. 血漿(jiang)：應根據標本的(de)要求選(xuan)擇EDTA或檸檬酸鈉作(zuo)為抗凝劑，混合(he)10-20分鐘后，離心(xin)20分鐘左右(you)（2000-3000轉(zhuan)/分）。仔細收集上清，保存過程中如有沉淀形成，應該再次離心(xin)。
3. 尿(niao)液：用無(wu)菌管收集(ji)，離心20分鐘左右（2000-3000轉/分）。仔細收集(ji)上清，保存過程中(zhong)如有(you)沉淀(dian)形成，應再次離心。胸腹水、腦脊液參(can)照實行。
4. 細(xi)胞培養上清(qing)(qing)：檢測分(fen)(fen)泌(mi)性的成(cheng)(cheng)份時，用無菌(jun)管(guan)收集(ji)(ji)。離(li)心(xin)20分(fen)(fen)鐘(zhong)左右(you)(you)（2000-3000轉(zhuan)/分(fen)(fen)）。仔(zi)細(xi)收集(ji)(ji)上清(qing)(qing)。檢測細(xi)胞內(nei)的成(cheng)(cheng)份時，用PBS（PH7.2-7.4）稀釋細(xi)胞懸(xuan)液，細(xi)胞濃度(du)達到100萬(wan)/ml左右(you)(you)。通過(guo)反復凍融，以使(shi)細(xi)胞破(po)壞并放出細(xi)胞內(nei)成(cheng)(cheng)份。離(li)心(xin)20分(fen)(fen)鐘(zhong)左右(you)(you)（2000-3000轉(zhuan)/分(fen)(fen)）。仔(zi)細(xi)收集(ji)(ji)上清(qing)(qing)。保存(cun)過(guo)程中如有(you)沉淀形成(cheng)(cheng)，應再次離(li)心(xin)。
5. 組織標(biao)本(ben)：切(qie)割標(biao)本(ben)后(hou)，稱取重量。加入一(yi)定量的(de)PBS，PH7.4。用液(ye)氮迅速(su)冷凍保存(cun)備用。標(biao)本(ben)融化后(hou)仍然保持2-8℃的(de)溫(wen)度。加入一(yi)定量的(de)PBS（PH7.4），用手工(gong)或(huo)勻漿(jiang)器將標(biao)本(ben)勻漿(jiang)充分(fen)。離心20分(fen)鐘左右（2000-3000轉/分(fen)）。仔細收集上清。分(fen)裝后(hou)一(yi)份待檢測，其余(yu)冷凍備用。
6. 標(biao)本采(cai)集(ji)后盡早進(jin)行(xing)提取(qu)，提取(qu)按相關文獻進(jin)行(xing)，提取(qu)后應(ying)盡快進(jin)行(xing)實驗(yan)。若(ruo)不(bu)能馬上進(jin)行(xing)試驗(yan)，可(ke)將標(biao)本放于-20℃保存，但應(ying)避免反復凍融.
7. 不能檢測含NaN3的樣品，因(yin)NaN3抑(yi)制辣根過氧化(hua)物酶的（HRP）活性。