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Hieff Unicon^? TaqMan multiplex qPCR master mix

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產(chǎn)品描述

本產(chǎn)品是2×Mix預(yù)混合試劑，能夠?qū)崿F(xiàn)在單個(gè)反應(yīng)孔中進(jìn)行多重?zé)晒舛?/span>PCR。本產(chǎn)品含有基因改造的熱啟動(dòng)Taq酶，極大地提高了擴(kuò)增靈敏度和特異性，本產(chǎn)品對(duì)多重反應(yīng)緩沖體系進(jìn)行了深度優(yōu)化，能夠提高反應(yīng)的擴(kuò)增效率，促進(jìn)低濃度模板的有效擴(kuò)增。本產(chǎn)品可用于基因分型和基因多重定量分析。

產(chǎn)品組分

組分編號(hào)	組分名稱	產(chǎn)品編號(hào)/規(guī)格
組分編號(hào)	組分名稱	11208ES03 (1 mL)	11208ES06 (5 × 1mL)	11208ES08 (5 mL)	11208ES60 (20×5 mL)
11208-A	2×Hieff Unicon^® TaqMan Multiplex qPCR Master Mix	1 mL	5 × 1 mL	5 mL	20 × 5 mL
11208-B	50×Low Rox	40 μL	200 μL	200 μL	20 × 200 μL
11208-C	50×High Rox	40 μL	200 μL	200 μL	20 × 200 μL

運(yùn)輸和保存方法

冰袋運(yùn)輸。-20°C儲(chǔ)存，有效期18個(gè)月。

反應(yīng)體系

組分	體積（μL）	終濃度
Hieff Unicon^® TaqMan Multiplex qPCR Master Mix	10	1×
Primer mix	x	0.1-1 μM
TaqMan Probe mix	y	50-300 nM
50×Low or High Rox	0.4	1×
模板DNA/cDNA	1-100 ng	-
ddH₂O	up to 20	-

【注】

a. 上表中模板量和引物濃度以及探針濃度均為推薦量，可根據(jù)具體實(shí)驗(yàn)情況進(jìn)行調(diào)整，選擇最適投入量和濃度

b. 一般情況下，每條引物濃度為 0.2 μM 即可得到較好的擴(kuò)增效果，引物濃度可在 0.1-1 μM 的范圍內(nèi)進(jìn)行調(diào)節(jié)；

c. 反應(yīng)體系可根據(jù)實(shí)驗(yàn)需求進(jìn)行等比例調(diào)節(jié)，模板體積不超過總體積的 1/10。

適用機(jī)型

Bio-Rad: CFX96, CFX384, iCycler iQ, iQ5, MyiQ, MiniOpticon, Opticon, Opticon 2, Chromo4;

Eppendorf: Mastercycler ep realplex, realplex 2 s;

Qiagen: Corbett Rotor-Gene Q, Rotor-Gene 3000, Rotor-Gene 6000;

Roche Applied Science: LightCycler 480, LightCycler 2.0; Lightcycler 96;

Thermo Scientific: PikoReal Cycler; Cepheid: SmartCycler; Illumina: Eco qPCR.

參考擴(kuò)增程序

循環(huán)步驟	溫度	時(shí)間	循環(huán)數(shù)
預(yù)變性	95°C	5 min	1
變性	95°C	5 sec	40
退火/延伸	60°C	30 sec	40

【注】

a. 某些情況下，可通過調(diào)節(jié)退火延伸時(shí)間提高多重 qPCR 反應(yīng)的性能；

b. 反應(yīng)結(jié)束后可通過瓊脂糖凝膠電泳確認(rèn)產(chǎn)物的特異性；

注意事項(xiàng)

1. 請(qǐng)于使用前確保產(chǎn)品完全解凍并徹底混勻，短暫離心收集至管底后置于冰上備用。

2. 為避免樣品間交叉污染和氣溶膠污染，推薦在超凈臺(tái)中進(jìn)行反應(yīng)體系的配制。

3. 使用過程中避免產(chǎn)品的反復(fù)凍融。

4. 為了您的安全和健康，請(qǐng)穿實(shí)驗(yàn)服并佩戴一次性手套操作。

HB220808

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[10] Song H, Xu Y, Shi L, et al. LncRNA THOR increases the stemness of gastric cancer cells via enhancing SOX9 mRNA stability. Biomed Pharmacother. 2018;108:338-346. doi:10.1016/j.biopha.2018.09.057(IF:3.457)
[11] Lou W, Yan J, Wang W. Downregulation of miR-497-5p Improves Sepsis-Induced Acute Lung Injury by Targeting IL2RB. Biomed Res Int. 2021;2021:6624702. Published 2021 Apr 12. doi:10.1155/2021/6624702(IF:3.411)
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