Infinizo USA SB 1000 упаковка из 2 анаэробных бактериальных септических емкостей очиститель и жидкости 1
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Стабильное качество продукта одноэтапный комплект rt qpcr (зонд) реагенты высокой чистоты
- Категория: Enzymes >>>
- Поставщик: Shijiazhuang Saidi Biotech Co. Ltd.
Сохранить в закладки 1600338531719:
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Характеристики
Product Description
Product name | One Step RT-qPCR Kit (Probe) |
Brand | SANSHIBIO |
Model | 50T |
MOQ | 200 |
One Step RT-qPCR Kit (Probe)
Product description
One Step RT-qPCR Kit (Probe) provides a convenient format for highly sensitive and specific RT-PCR using any RNA template. The kit includes optimized components that allow both reverse transcription and PCR amplification to take place in the same reaction mix in a "one-step" reaction, which is simple to operate, avoids cross-contamination between samples and improves the sensitivity of detection. One Step RT-qPCR Enzyme Mix of the kit contains HI-MMLV Reverse Transcriptase, antibody-modified hot-start Taq DNA polymerase and RNase Inhibitor, which has stronger RNA affinity and thermal stability, as well as higher amplification efficiency and specificity. The kit can obtain a good standard curve in a wide quantitative area, and accurately and quantitatively detect a variety of high and low abundance target genes, with good repeatability and high reliability.Product content
One Step RT-qPCR Enzyme Mix, 5x One Step RT-qPCR Master Mix, RNase-Free ddH2O
Operation method
1. Prepare the following reaction solutions in RNase-Free centrifuge tubes.
Reagent Volume
One Step RT-qPCR Enzyme Mix 1µl
5x One Step RT-qPCR Master Mix 4µl
Forward Primer (10µM) 0.4µl
Reverse Primer (10µM) 0.4µl
Probe (10µM) 0.2µl
Total RNA 1pg-1µg
RNase-Free ddH2O Up to 20µl
The amount of each component in the reaction system can be adjusted according to the following principles:
1) Generally speaking, the final concentration of primers in the reaction system is 0.2 μM to get a better amplification effect. When the reaction performance is poor, the primer concentration can be adjusted within the range of 0.1-1.0 µM.
2) The probe concentration can be adjusted between 50-250 nM.
3) The qPCR is extremely flexible. The accuracy of the amount of template added when setting up the reaction system has a great impact on the final quantitative results. It is recommended to add the template diluted (such as diluting 2-5μl sample) into the reaction system, which can be effective improve the repeatability of the experiment.
4) Please choose the length of the amplified product within the range of 80-300 bp.
5) If using ABI series Real Time PCR instrument, add 50x ROX reference dye when preparing the solution.
2. Perform the One Step RT-qPCR reaction according to the following conditions.
Method/Step\tStandard procedure\tFast procedure\tNumber of cycles
50℃\t 15min\t 10min\t 1
95℃\t 3min\t 1min\t 1
95℃\t 15s\t 5s\t 45Cycles
60℃\t 30s\t 20s\t 45Cycles
a. For templates with complex secondary structures and high GC regions, set the reverse transcription temperature to 55°C, which helps to improve amplification efficiency and sensitivity.
b. Please adjust the extension time according to the minimum time limit of data collection required by the Real Time PCR instrument used. At least 30s when using ABI 7700 and ABI 7900, at least 31s when using ABI 7000 and ABI 7300, and at least 34s when using ABI 7500.
c. Whether the Real Time PCR instrument actually used supports rapid amplification cycle, please perform pre-experiment.
3. Analysis of experimental results:
After the reaction, the experimental results are analyzed. For detailed analysis methods, please refer to the Real Time PCR Amplifier Operation Manual.
Precautions:
1) One Step RT-qPCR Enzyme Mix contains a high concentration of glycerol. Please centrifuge briefly before using, collect it to the bottom of the reaction tube, and gently blow with a pipette to mix well before using.
2) Please use RNase-Free pipette tips, EP tubes, etc. to prepare the reaction solution to avoid contamination as much as possible.
3) The qPCR is extremely flexible. The accuracy of the amount of template added when setting up the reaction system has a great impact on the final quantitative results. It is recommended to add the template diluted (such as diluting 2-5μl sample) into the reaction system, which can be effective improve the repeatability of the experiment.
4) Please choose the length of the amplified product within the range of 80-300 bp.
5) If using ABI series Real Time PCR instrument, add 50x ROX reference dye when preparing the solution.
2. Perform the One Step RT-qPCR reaction according to the following conditions.
Method/Step\tStandard procedure\tFast procedure\tNumber of cycles
50℃\t 15min\t 10min\t 1
95℃\t 3min\t 1min\t 1
95℃\t 15s\t 5s\t 45Cycles
60℃\t 30s\t 20s\t 45Cycles
a. For templates with complex secondary structures and high GC regions, set the reverse transcription temperature to 55°C, which helps to improve amplification efficiency and sensitivity.
b. Please adjust the extension time according to the minimum time limit of data collection required by the Real Time PCR instrument used. At least 30s when using ABI 7700 and ABI 7900, at least 31s when using ABI 7000 and ABI 7300, and at least 34s when using ABI 7500.
c. Whether the Real Time PCR instrument actually used supports rapid amplification cycle, please perform pre-experiment.
3. Analysis of experimental results:
After the reaction, the experimental results are analyzed. For detailed analysis methods, please refer to the Real Time PCR Amplifier Operation Manual.
Precautions:
1) One Step RT-qPCR Enzyme Mix contains a high concentration of glycerol. Please centrifuge briefly before using, collect it to the bottom of the reaction tube, and gently blow with a pipette to mix well before using.
2) Please use RNase-Free pipette tips, EP tubes, etc. to prepare the reaction solution to avoid contamination as much as possible.
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FAQ
Q1- Is the MOQ negotiable?
A1- Yes, MOQ is negotiable, while the unit price is different according to the order quantity.
Q2- How long is the lead time?
A2- Lead time depends on the order quantity. Normally 15-25 days and special case 40-50 days in sales peak season. Please contact us for details.
Q3- Can you make the according to my design?
A3- Yes, we can. Please contact us with your requirement or CAD drawing.
Q4- Can I arrange third party inspection?
A4- Yes, we welcome QC from customers and third-party inspection.
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