Tailing miRNA dye fluorescence quantitative kit

Tailing miRNA dye fluorescence quantitative kit

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Molecular Biology

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Tailing miRNA dye fluorescence quantitative kit Cat. No.: MR***1 Price: $*0 for *5T(negotiable) Storage temperature: **0. C, store away from light for at least 6 months, mix thoroughly before use Product components

components	MR***1
2x miRNA qPCR Mix(with SYBR Green)	1.*5ml
Reverse primer(*0uM)	*5ul

Product Description The miRNA Fluorescence Quantitative PCR Detection Kit uses the principle of SYBR Green I chimeric fluorescence method for miRNA fluorescence quantitative detection. This kit contains all reagents for miRNA fluorescence quantitative detection, including 2×miRNA qPCR Mix and Reverse primer. 2×miRNA qPCR Mix (containing Sybr Green) is a new generation of premixed fluorescence quantitative PCR detection reagents specially developed for miRNA quantitative detection. The DNA polymerase in it adopts the hot start form of antibody modification, and with a special buffer system, the reaction specificity is better, the sensitivity is higher, and accurate quantification can be performed in a wider range. Note: This kit must be used in conjunction with the miRNA First Chain Synthesis Kit (MF***1). Reagents to be prepared: 1. Molecular biology experimental grade water (nuclease-free) 2. qPCR upstream primer (Forward primer) corresponding to the miRNA to be detected Forward primer design principles 1. Follow the most common principles of primer design 2. Based on the mature miRNA sequence, replace U with T, 3. The Tm value of the downstream primer provided in the kit is *5. C 4. If the Tm value of the primer directly designed according to principle 2 is too low, you can add a few bases (preferably G or C bases) to the 5' end of the primer; you can also add one or more A bases to the 3' end; or modify both the 5' end and the 3' end; 5. If the Tm value of the primer directly designed according to principle 2 is too high, you can remove a few bases at the 5' end or 3' end of the primer. Notes: 1. The amount of miRNA first-chain cDNA added should not exceed 1/*0 of the volume of Real time PCR 2. For special detection systems, high-content cDNA templates are prone to non-specific amplification. Dilute cDNA appropriately (*0 times or **0 times) according to the abundance of the miRNA being detected 3. This product contains the fluorescent dye SYBR Green I. Avoid strong light exposure when storing this product or preparing PCR reaction solution. 4. 2 x miRNA qPCR Mix does not contain the reference dye ROX. Customers can decide whether to add ROX according to the instructions of the qPCR instrument to eliminate the signal background and correct the fluorescence signal error between wells. Operation steps 1. Melt 2x miRNA qPCR Mix (with SYBR Green) and Reverse primer (*0uM) at room temperature. 2. When using, turn the 2x miRNA qPCR Mix upside down and gently mix it evenly to avoid bubbles, and use it after slight centrifugation. If the reagents are not mixed well, the reaction performance will be reduced (please do not use an oscillator to mix). 3.Prepare the reaction solution on ice according to the components in the table below


reagents	Amount of usage		Final concentration
Taq SYBR® Green qPCR Premix	*5ul	*0ul	1×
Forward primer (*0uM)	1ul	0.4ul	0.2uM
Reverse primer(*0uM)	1ul	0.4ul	0.2uM
miRNA第一链cDNA	X ul	X ul	*-
Nuclease-Free Water	To *0ul	To *0ul

PCR cycle (three-step method)

Procedure	temperature	time
Initial denaturation	*4℃	**3min
denaturation	*4℃	****0 sec	****5个cycling
annealing	5~5℃	****0 sec
elongation	*2℃	****0 sec
Dissociation curve	Use the instrument default acquisition program

PCRcycle (two-step method)

Procedure	temperature	time
Initial denaturation	*4℃	**3min
denaturation	*4℃	****0 sec	****5个cycling
Annealing&elongation	*0℃	*0 sec	****5个cycling
Dissociation curve	Use the instrument default acquisition program

Note: To improve specificity, select the two-step method; to improve amplification efficiency, select the three-step method.

国家:	China
型号:	-
离岸价格:	（面議）获取最新报价
位置:	China
最小订单价格:	-
最小订单:	-
包装细节:	-
交货时间:	-
供应能力:	-
付款方式:	-
產品組 :	Molecular Biology