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BioTNT Realtime PCR mRNA primer pair引物对(全基…
    产品中心    >>  试剂    >>  分子生物学试剂    >>  qPCR kits & 核酸抽提 关   闭
Realtime PCR mRNA primer pair引物对(全基因组全种属)套装试剂盒(10对mRNA引物、1支 premix)
分类:qPCR kits & 核酸抽提
品牌:BioTNT
货号:PRIM10B1
原价:3310
现价:¥2549.00    节省:¥761
规格:1 kit
说明:10对mRNA引物、1支?premix
产品参数

 

配套试剂
RNA 抽提试剂盒 逆转录试剂盒 Real time PCR 试剂盒 Premix

 

订购指南
货 号 组 成 250T 500T
PRIM013475 1对mRNA引物 ¥299 ¥399
PRIM013475B1 1对mRNA引物、1支 premix ¥549 ¥649
PRIM013475B5 1对mRNA引物、5支 premix ¥1469 ¥1569
PRIM013475B10 1对mRNA引物、10支 premix ¥2399 ¥2499
PRIM013475C 1对mRNA引物、1对内参 ¥369 ¥469
PRIM013475C1 1对mRNA引物、1对内参和1支 premix ¥619 ¥719
PRIM013475C5 1对mRNA引物、1对内参和5支 premix ¥1519 ¥1619
PRIM013475C10 1对mRNA引物、1对内参和10支 premix ¥2449 ¥2549
PRIM5 5对mRNA引物 ¥1299 ¥1799
PRIM5B1 5对mRNA引物、1支 premix ¥1549 ¥2049
PRIM5B5 5对mRNA引物、5支 premix ¥2469 ¥2969
PRIM5B10 5对mRNA引物、10支 premix ¥3399 ¥3899
PRIM5C 5对mRNA引物、1对内参 ¥1369 ¥1869
PRIM5C1 5对mRNA引物、1对内参和1支 premix ¥1619 ¥2119
PRIM5C5 5对mRNA引物、1对内参和5支 premix ¥2519 ¥3019
PRIM5C10 5对mRNA引物、1对内参和10支 premix ¥3449 ¥3949
PRIM10 10对mRNA引物 ¥2299 ¥3299
PRIM10B1 10对mRNA引物、1支 premix ¥2549 ¥3549
PRIM10B5 10对mRNA引物、5支 premix ¥3469 ¥4469
PRIM10B10 10对mRNA引物、10支 premix ¥4399 ¥5399
PRIM10C 10对mRNA引物、1对内参 ¥2369 ¥3369
PRIM10C1 10对mRNA引物、1对内参和1支 premix ¥2619 ¥3619

 

 

使用说明
PRINCIPLE OF THE ASSAY

RT-QPCR Primer Assays are designed for SYBR Green based on real-time PCR detection. The primer design computer algorithm has been developed using an in vitro assay to ensure that the resulting primer sequences generate a single PCR product of the predicted size and a minimal amount of primer–dimer in 30 cycles of PCR amplification. The assay also ensures that the amplification efficiency of the primers is at least 90%. As a result, the algorithm designs highly effective primer sequences for SYBR Green based real-time PCR detection. RT-QPCR Primer Assays are available for every human, mouse, rat, rhesus macaque, fruit fly, and dog gene annotated by the NCBI.

Advantage:

Uniform Reaction Condition: Reduce experimental design time

High Specificity: Absence of non-specific amplification and primer-dimers ensures reproducible data

Validation and Precision: 
a) Human-,mouse-,rat-and rabbit specific primers are designed by a proprietary algorithm and validated for precision performance. 
b) Primer validation includes melting curve to ensure amplification of the correct target DNA.

Component Volume Storage
Biotnt qPCR PreMix 400 uL -20℃
Upper Primer 1 000 uL -20℃
Lower Primer 1 000 uL -20℃
nuclease-free H2O 1 000 uL -20℃

*Store Nuclease-free Water at -20℃、4℃ or room temp.

qPCR Detection Kit is shipped at ambient temperature, But for long time storage, RT-QPCR Detection kit can be used within 12 mouths if the whole kit is stored at -20℃


i. Briefly (10-15 seconds) spin down all reagents.

ii. For each 20µl PCR, mix the following components in a PCR tube.
Per reaction Component
Real time PCR Premix 10µl
Upper Primer 2µl
Lower Primer 2µl
cDNA 1µl
Nuclease-free Water To 20µl

Note: We suggest adjusting RNA concentration so that 1 μL is used

iii. Quickly centrifuge and place your tubes in your real-time thermal cycler. iv. Enter and run the appropriate program for your real-time instrument. 

qPCR system setup

a) Use a two-step cycling program, for the following instrumentation.
PCR system Cycle Time Temp

ABI:, Vii7,7700,7900,StepOnePlus,

Bio-Rad:iCycler®, iQ5, MyIQ

Stratagene: Mx3000p, Mx3005p, Mx4000p

Eppendorf: Mastercycler® ep realplex

1 5 minutes [1] 95℃
40 15 seconds [2] 95℃
30 seconds [3] 60℃
Roche*: LightCycler 480® 1 10minutes 95℃
45 15seconds 95℃
1minute 60℃

Attention: Roche LightCycler 480® Users: Adjust the ramp rate to 1ºC/sec.


b) Use a three-step cycling program, for the following instrumentation
PCR system Cycle Time Temp

ABI:Vii7,StepOnePlus,7700,7900,

Bio-Rad:iCycler®, iQ5, MyIQ

Stratagene: Mx3000p, Mx3005p, Mx4000p

Eppendorf: Mastercycler® ep realplex

1 10 minutes [1] 95℃
45 15 seconds [2] 95℃
30-40seconds 60℃
30 seconds[3]

1. The 10-minute step at 95ºC is required to activate the HotStart DNA polymerase.

2. Detect and record SYBR Green fluorescence from every well during the annealing step of each cycle.

3. Different instruments need different lengths of time to detect the fluorescent signal. Choose the appropriate time for the annealing step (55ºC) for your instrument.




Reference:

1. Xi Chen1,Zheng-Yun Zhang2. Characterization of mesenchymal stem cells under the stimulation of Toll-like receptor agonists[J]. Development, Growth & Differentiation,2014,5(2): 233–244. DOI: 10.1111/dgd.12124

2. Xiaoling Tao1,Jinqi Fan1,Guoying Kao1. Angiotensin-(1–7) attenuates angiotensin II-induced signaling associated with activation of a tyrosine phosphatase in Sprague-Dawley rats cardiac fibroblasts[J]. Biology of the Cell, Accepted Article. DOI: 10.1111/boc.201400015

3. Xuelian Chen MD,Yan Liu MD. Topical insulin application improves healing by regulating the wound inflammatory response[J]. Wound Repair and Regeneration,2012,20(3): 425–434. DOI: 10.1111/j.1524-475X.2012.00792.x

4. Jiawen Fan1,Gezhi Xu1,2 . Pharmacologic Induction of Heme Oxygenase-1 Plays a Protective Role in Diabetic Retinopathy in Rats[J].Investigative ophthalmonogy & visual science,2012,(53): 6541-6556. doi: 10.1167/iovs.11-9241

5. Xiangyu Zou1,Guangyuan Zhang1. Microvesicles derived from human Whartons Jelly mesenchymal stromal cells ameliorate renal ischemia-reperfusion injury in rats by suppressing CX3CL1[J]. Stem Cell Research & Therapy,2014,(5):40. doi:10.1186/scrt428

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