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产品中心 >> 试剂 >> 分子生物学试剂 >> qPCR kits & 核酸抽提
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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、10支 premix)
分类:qPCR kits & 核酸抽提
品牌:BioTNT
货号:PRIM10B10
原价:¥5720
现价:¥4399.00 节省:¥1321
规格:1 kit 说明:10对mRNA引物、10支?premix
产品参数
配套试剂
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℃
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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.
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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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