Description
Hieff Unicon™ Lyo-Ready I TaqMan Multiplex qPCR Kit is a fluorescence quantitative qPCR lyophilization reagent using the heat start Taq enzyme using a new generation antibody method, Suitable for FAM / HEX / TET / JOE / ROX double probe (this product is also suitable for Molecular Beacon probe), Reagents contain components including lyophilized protectants, heat start Fast Taq DNA polymerase, dNTP, Can be directly lyophilized on the machine. This product does not contain ROX correction dye, suitable to various Quantitative PCR models.
Specifications
|
Hot Start |
Built-in hot start |
|
Detection method |
Primer-probe detection |
|
PCR method |
qPCR |
|
Polymerase |
Taq DNA polymerase |
|
Type of sample |
DNA |
|
Application equipment |
Applied Biosystems: 5700, 7000, 7300, 7700, 7900HT Fast, StepOne™, StepOne Plus™, 7500, 7500 Fast, ViiA™7, QuantStudio™ 3 and 5, QuantStudio™ 6,7,12k Flex; Bio-Rad: CFX96, CFX384, iCycler iQ, iQ5, MyiQ, MiniOpticon, Opticon, Opticon 2, Chromo4; Eppendorf: Mastercycler ep realplex, realplex 2 s; Qiagen: Corbett Rotor-Gene Q, Rotor-Gene 3000, Rotor-Gene 6000; Roche Applied Science: LightCycler 480, LightCycler 2.0; Lightcycler 96; Stratagene: MX3000P™, MX3005P™, MX4000P™; Thermo Scientific: PikoReal Cycler; Cepheid: SmartCycler; Illumina: Eco qPCR; SLAN:SLAN-96S,SLAN-96P.. |
Components
Storage
The product is shipped with dry ice and can be stored at -15℃ ~ -25℃ for 1 year. .
Instructions
- Lyophilized and machine loading system
|
Components |
Volume (μL) |
Final Concentration |
|
Lyo-ReadyTaqMan qPCR Mix |
20 |
1× |
|
Primer mix (10 μmol/L) |
x |
0.1-0.5 μmol/L |
|
Probe mix (10 μmol/L) |
x |
50-250 nmol/L |
|
Total Volume |
21-25 |
- |
*The primer concentration can be adjusted appropriately according to the experimental requirements, and the total volume should not exceed 5 μL.
- Lyophilization procedure
|
Step |
Temperature |
Set time |
Duration |
Vacuum degree |
|
Pre-Freezing |
-45℃ |
10 min |
60 min |
—— |
|
One-time dry stage |
-45 ℃ |
10min |
60min |
0.01mbar |
|
-40 ℃ |
10min |
600min |
0.01mbar |
|
|
-35 ℃ |
10min |
120min |
0.01mbar |
|
|
-30 ℃ |
20min |
60min |
0.01mbar |
|
|
-20 ℃ |
20min |
60min |
0.01mbar |
|
|
-10 ℃ |
20min |
60min |
0.01mbar |
|
|
0 ℃ |
20min |
60min |
0.01mbar |
|
|
Pressure Rise Test |
||||
|
Analyze the drying stage |
5 ℃ |
10min |
20min |
0.01mbar |
|
10 ℃ |
10min |
20min |
0.01mbar |
|
|
15 ℃ |
10min |
20min |
0.01mbar |
|
|
20 ℃ |
10min |
20min |
0.01mbar |
|
|
25 ℃ |
10min |
240min |
0.01mbar |
|
|
Pressure Rise Test |
||||
- Reaction System
|
Components |
Volume (μL) |
Final Concentration |
|
20 μL Lyo-Bead |
1 bead |
/ |
|
Primer mix (10 μmol/L) |
x |
0.1-0.5 μmol/L |
|
Probe mix (10 μmol/L) |
x |
50-250 nmol/L |
|
Template |
1-25 |
1× |
|
ddH2O |
up to 25 |
- |
*The template is dissolved in ddH2O, and the elution solution of the nucleic acid extraction reagent is using ddH2O.
- Reaction program
|
Cycle step |
Temp. |
Time |
Cycles |
|
Initial denaturation |
95℃ |
5 min |
1 |
|
Denaturation |
95℃ |
15 s |
45 |
|
Annealing/Extension |
60℃ |
30 s |
*The annealing / extension temperature can be adjusted appropriately according to the experimental requirements.
Notes
Please wear the necessary PPE, such lab coat and gloves, to ensure your health and safety！