One Step RT-qPCR Probe Kit
Cat：P***2 Price: $**4 for **0T
Storage conditions: Store at **0℃; avoid
repeated freezing and thawing.
Product Introduction
One Step RT-qPCR Probe Kit v2 is a kit for
multiple reverse transcription-fluorescence quantitative PCR
reactions using RNA as a template. During the experiment, reverse
transcription and quantitative PCR are performed in the same
reaction tube, which simplifies the experimental operation and
reduces the risk of contamination. This kit uses heat-resistant
Reverse Transcriptase to efficiently synthesize the first-strand
cDNA, and uses hot-start Ab-Taq DNA Polymerase for quantitative
amplification, which is suitable for probe-based qPCR. This product
contains all reverse transcription-fluorescence quantitative PCR
components except primers and sample RNA, including RNase
inhibitors, RTase, and Ab-Taq. It can reduce the number of
operation steps, shorten the sample addition time, and reduce the
chance of contamination. Compatible models
Rox calibration is not required: Bio-Rad full
series, Roche full series, Eppendorf full series, Takara full
series, Qiagen full series, Hongshi full series, Tianlong full
series, Biori full series, Yarui full series,
etc. High Rox needs to be added: ABI ***0, ***0,
***0, ***0, ***0HT Fast, StepOne™, StepOnePlus™, etc.; Low Rox
needs to be added: ABI ***0, ***0 Fast, ViiA™7, QuantStudio™ 3 and
5, tratageneMX***0P™, MX***5P™, MX***0P™, etc.
Instructions
1. Precautions for use
â‘ Before use, it should be fully thawed at
room temperature, mixed and centrifuged
instantly; â‘¡ Avoid bubbles in the reaction solution as
much as possible; â‘¢ The RNA to be tested should be as fresh as
possible, and RNase contamination should be strictly prevented
during the extraction process.
|
|
|
|
Reagents |
Amount of Usage |
Final concentration |
|
RTQ Enzyme Mix |
1 μl |
/ |
|
2×RTQ Reaction Buffer |
*0 μl |
/ |
|
forward primer
(*0 μM)a |
0.4 μl |
0.2 μM |
|
reverse
primer (*0 μM)a |
0.4 μl |
0.2 μM |
|
TaqMan 探针 (5 μM)b |
1 μl |
0.*5 μM |
|
RNA 模æ¿c |
1 pg~1 μg |
/ |
|
Nuclease-Free Water |
To *0 μl |
|
a. The recommended final concentration of
primers is 0.2 μM. If the effect is not good, it can be adjusted to
0.1~1 μM; please set the primer length to *8~*5 bp and the GC
content to *0%~*0%; b. The recommended final concentration of probes
is 0.*5 μM. If the effect is not good, it can be adjusted to 0.1~1
μM; c. The recommended template loading volume is
1~5 μl. qPCR has extremely high sensitivity. It is recommended to
dilute the template and control the Ct value between
*0~*5; d. Please prepare in a clean bench and use a gun
tip and reaction tube without nuclease residue; it is recommended
to use a gun tip with a filter. Avoid cross contamination and
aerosol contamination. 2. Reaction procedure (can be adjusted
appropriately according to the model)
|
procedure |
temperature |
time |
|
|
reverse transcription |
*0℃ |
*0min |
|
|
predegeneration |
*7℃ |
1 min |
|
|
denaturation |
*7℃ |
5 sec |
*0~*5
cycling |
|
Annealing &
Extension a |
*8℃ |
*0 sec |
Notes Please use RNase free consumables during the
experiment.
