乙型流感診斷試劑盒（快檢方法）

廣州健侖生物科技有限公司

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乙型流感診斷試劑盒（快檢方法）

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【公司名稱】 廣州健侖生物科技有限公司
【市場部】    楊永漢

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【騰訊  】 2042552662
【公司地址】 廣州清華科技園創(chuàng)新基地番禺石樓鎮(zhèn)創(chuàng)啟路63號(hào)二期2幢101-103室

實(shí)驗(yàn)設(shè)備
1. 電泳設(shè)備
2. 紫外燈
實(shí)驗(yàn)步驟
1．電泳槽用0.3%H2O2浸泡30min，DEPC水沖洗晾干。 
2．鋪膠（40ml） 
       瓊脂糖                0.55g   
       DEPC水             36ml 
      10×MOPS         5ml 
      37%甲醛            9ml 
稱0.55克瓊脂糖加36ml DEPC水，微波爐加熱溶解瓊脂糖，肉眼觀察無顆粒狀懸浮物。冷卻至50-60℃，加9ml 甲醛（37%）和5ml 10×電泳緩沖液，然后灌制凝膠，插入合適長度和寬度的梳子，于室溫放置30分鐘以上使凝膠凝固。
3．RNA樣品處理（以一條泳道為例）一般取0.3 g的總RNA，加入1/5體積的5×加樣緩沖液，65℃加熱5 min，冰上驟冷，以消除RNA的二級(jí)結(jié)構(gòu)。建議上樣前在RNA樣品中加入0.5-1.0 ul的溴化乙錠（EB，濃度1.0 mg/mL），而不在膠中加EB，這樣電泳后的背景較低。配好的甲醛變性膠先在1×甲醛變性膠電泳緩沖液中預(yù)電泳15min。RNA樣品在5-10 V/cm的電壓降下電泳30min。
4．加2.0ml甲醛凝膠加樣緩沖液（10×） 
5．加樣和電泳 
將凝膠預(yù)電泳15min，電壓降為5-10 v/cm。隨后加樣品和標(biāo)準(zhǔn)物，以3-4v/cm電壓降電泳，電泳液為1×MOPS電泳緩沖液。直至溴酚藍(lán)遷移至膠下游的3/4處。

Laboratory equipment
Electrophoresis equipment
2. UV lamp
Experimental steps
1. Electrophoresis tank with 0.3% H2O2 soak 30min, DEPC water rinse to dry.
2. Shop plastic (40ml)
       Sepharose 0.55g
       DEPC water 36ml
      10 × MOPS 5ml
      37% formaldehyde 9ml
Weigh 0.55 grams of agarose and 36 ml of DEPC water, dissolve the agarose in a microwave oven, and visually observe the non-granular suspension. Cool to 50-60 ° C. Add 9ml of formaldehyde (37%) and 5ml of 10x running buffer then gel and insert a comb of appropriate length and width for 30 minutes at room temperature to allow the gel to set.
3. RNA sample processing (taking one lane as an example) Generally, take 0.3 g of total RNA, add 1/5 volume of 5 × loading buffer, heat at 65 ° C for 5 min and quench on ice to eliminate RNA secondary structure. It is recommended that 0.5-1.0 μl of ethidium bromide (EB, 1.0 mg / mL) be added to the RNA sample prior to loading without adding EB to the gel so that the background after electrophoresis is low. Formaldehyde denatured plastic first in 1 × formalin gel electrophoresis buffer pre-electrophoresis 15min. RNA samples were electrophoresed for 30 min at a voltage drop of 5-10 V / cm.
4. Add 2.0ml formaldehyde gel loading buffer (10 ×)
5. Sample and electrophoresis
The gel pre-electrophoresis 15min, the voltage drop to 5-10 v / cm. Then add samples and standards, with 3-4v / cm voltage drop electrophoresis, electrophoresis solution 1 * MOPS electrophoresis buffer. Until bromophenol blue migrates to 3/4 of the gel downstream.