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One tube multiplex for detection of West Nile virus and internal control

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西尼羅河病毒檢測(cè)試劑盒（PCR-熒光探針?lè)ǎ?/strong>

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【公司名稱】 廣州健侖生物科技有限公司
【市場(chǎng)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

一、基本知識(shí)與原理 
轉(zhuǎn)導(dǎo)是以噬菌體為媒介將一個(gè)細(xì)胞的遺傳物質(zhì)轉(zhuǎn)移給另一個(gè)細(xì)胞的過(guò)程。隨著分子遺傳學(xué)的發(fā)展，轉(zhuǎn)身已成為基因精細(xì)結(jié)構(gòu)分析的常用方法之一。 
根據(jù)噬菌體轉(zhuǎn)導(dǎo)供體菌基因的差異，轉(zhuǎn)導(dǎo)可分為普遍性轉(zhuǎn)導(dǎo)和局限性轉(zhuǎn)導(dǎo)。這里以局限性轉(zhuǎn)導(dǎo)為例說(shuō)明轉(zhuǎn)導(dǎo)的基本原理。局限性轉(zhuǎn)導(dǎo)實(shí)驗(yàn)中常用的是大噬菌體，它能整合在大腸桿菌染色體DNA上的半乳糖基因（gAl）和生物素基因（bib）之間，因此，它能轉(zhuǎn)導(dǎo)半乳糖基因（一）又能轉(zhuǎn)導(dǎo)生物素基因（bio）。 
本實(shí)驗(yàn)選用大腸桿菌 E。oh K．2（A）為供體菌（即大噬菌體的DNA已整合在大腸桿菌的DNA上，我們稱該大腸桿菌為溶源性大腸桿菌，’gAT””為帶有半乳糖基因）。由于在此供體菌中噬菌體與半乳糖基因（匆”）緊密連鎖，因此，當(dāng)此供體菌受紫外線照射后會(huì)產(chǎn)生裂解反應(yīng)，噬菌體被誘發(fā)釋放，以一定的比例形成帶有半乳糖基因的轉(zhuǎn)導(dǎo)噬菌體。當(dāng)這種轉(zhuǎn)導(dǎo)噬菌體與受體菌 ECo］i K；。 s gAl－（此細(xì)菌不能利用半乳糖，‘’表示此細(xì)菌半乳糖基因發(fā)生突變）混合接觸時(shí)，帶有一基因的轉(zhuǎn)導(dǎo)噬菌體能以一定的頻率整合到受體首上，從而使不能利用半乳糖的 gAl一受體菌轉(zhuǎn)變成了能利用半乳糖的 gAT”細(xì)菌。整個(gè)過(guò)程可用圖12－’表示：l 
二、實(shí)驗(yàn)?zāi)康暮鸵?nbsp;
以局限性轉(zhuǎn)導(dǎo)為例來(lái)說(shuō)明轉(zhuǎn)導(dǎo)的基本原理，進(jìn)一步驗(yàn)證是遺傳物質(zhì)，并初步掌握轉(zhuǎn)導(dǎo)實(shí)驗(yàn)的基本方法。 
三、實(shí)驗(yàn)器材 
（1）實(shí)驗(yàn)材料：供體菌 受體菌 
（2）實(shí)驗(yàn)試劑：肉湯液體培養(yǎng)基、肉湯固體培養(yǎng)基、ZE 肉湯液體培養(yǎng)基、半固體瓊脂培養(yǎng)基、半乳糖Emb培養(yǎng)基、滅菌生理鹽水（或磷酸緩沖液） 
D（3）實(shí)驗(yàn)設(shè)備：培養(yǎng)皿（9厘米）、三角燒瓶（50毫升）、試管、離心管，離心機(jī)，移液管，涂棒，水浴鍋，離心機(jī)，紫外照射箱、溫箱 

First, the basic knowledge and principles
Transduction is the process of transferring a cell's genetic material to another cell using phage as a medium. With the development of molecular genetics, turning has become one of the commonly used methods for gene fine structure analysis.
According to the phage transduction donor gene differences, transduction can be divided into universal transduction and localized transduction. Here is a case of limited transduction as an example of the basic principle of transduction. Commonly used in limited transduction experiments is the large phage, which integrates between the galactose gene (gAl) and the biotin gene (bib) on the chromosomal DNA of E. coli so that it can transduce the galactose gene (a) But also biotin gene (bio).
E. coli E used in this experiment. oh K. 2 (A) is the donor bacteria (ie, the DNA of the phage is integrated into the DNA of E. coli, which we call lysogenic E. coli and the "gAT" is the galactose-bearing gene). Since the bacteriophages are closely linked to the galactose gene (hASH) in this donor bacterium, when the donor bacterium is lysed by UV rays, a cleavage reaction occurs and the phage are induced to release, forming a certain ratio of the galactose gene Of the transduced phage. When this transduced phage and recipient bacteria ECo] i K ;. s gAl- (the bacteria can not use galactose, '' means that the bacterial galactose gene mutation) mixed contact with a gene Of the transductant phage can be integrated into the receptor at a certain frequency on the first, so that can not use galactose gAl a receptor bacteria into galactose gAT "bacteria. The whole process can be shown in Figure 12- ': l
Second, the purpose and requirements of the experiment
The limitations of transduction as an example to illustrate the basic principles of transduction, further validation of genetic material, and preliminary grasp of the basic methods of transduction experiments.
Third, experimental equipment
(1) Experimental material: donor bacteria bacteria
(2) Experimental Reagents: broth broth, broth solid broth, ZE broth broth, semi-solid agar broth, galactose Emb medium, sterile saline (or phosphate buffer)
(3) Experimental equipment: Petri dishes (9 cm), Erlenmeyer flask (50 ml), test tube, centrifuge tube, centrifuge, pipette, dip stick, water bath, centrifuge, UV irradiation box,