SuperFast RT Master Mix for qPCR (gDNA remover) |
| カタログ番号GK10031 |
SuperFast RT Master Mix for qPCR (gDNA remover) is a reverse transcription kit with ultra-high sensitivity. 8-10 mins to complete the removal of gDNA and synthesis of cDNA.
Products are for research use only. Not for human use. We do not sell to patients.
Sample solution is provided at 25 µL, 10mM.
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SuperFast RT Master Mix for qPCR (gDNA remover) is a reverse transcription kit with ultra-high sensitivity. It consists of a reverse transcription reaction premix and a gDNA removal mix and also provides a portion of DEPC-ddH2O for dilution. Obtaining cDNA with SuperFast RT Master Mix for qPCR (gDNA remover) is the first step of a two-step RT-PCR experiment. The obtained cDNA can be used for conventional PCR or RT-PCR. If you are doing an RT-PCR experiment, it is recommended to use it with SYBR Green qPCR Master Mix (Catalog Number: GK10002).
SuperFast RT Master Mix for qPCR (gDNA remover) uses the high temperature (>50℃) high-speed amplification reverse transcriptase Super Reverse Transcriptase, which can synthesize cDNA from extremely low amounts (pg~μg level) of total RNA or poly(A) mRNA and is compatible with RNA templates with high GC content and complex secondary structure.
In many cases, RNA extracted by methods such as spin columns or acid guanidine-phenol chloroform (AGPC) method will contain a small amount of genomic DNA (gDNA). When pseudogenes exist in the target gene or primers cannot be designed across introns, genomic DNA will be amplified together with cDNA, thus affecting the accuracy of the data. SuperFast RT Master Mix for qPCR (gDNA remover) contains a gDNA remover with strong DNA degradation ability, which helps to remove gDNA and reverse transcribe RNA without purification.
Features of SuperFast RT Master Mix for qPCR
1. Effective removal of gDNA: RT Master Mix only takes 2 minutes to remove genomic DNA contamination.
2. High efficiency and fast: This kit uses extremely fast amplification reverse transcriptase and optimized oligonucleotide dT primers to achieve efficient reverse transcription, and the reaction can be completed in just 5 minutes.
3. Ultra-high sensitivity: Good reverse transcription reaction can also be performed on very small amounts of RNA templates.
4. Super compatibility: RT Master Mix can be compatible with RNA templates with high GC content and complex secondary structure.
5. Ultra-low impact factors: Super strong applicability to subsequent qPCR reactions: Through continuous optimization, the impact of this kit on subsequent qPCR reactions is minimized.
Operation flow:
1. Prepare the reaction system according to the table below (operate on ice), with a total volume of 10μl.
|
5x gDNA Remover Mix |
2μl |
|
RNA template |
0.01~1μg |
|
DEPC-ddH2O |
Add to 10μl |
Note:
(1) To ensure the accuracy of the reaction solution preparation, it is recommended to prepare the premixed system according to the amount of reaction number+2, and then dispense it into each reaction tube, and finally add RNA;
(2) If the amount of RNA to be used is greater than 1μg, it is recommended to scale up the reaction system proportionally.
2. Gently mix the reaction solution, centrifuge briefly to concentrate the solution at the bottom of the tube, incubate at 42℃ for 2 minutes, and then immediately place on ice to cool.
3. Add the following ingredients to the tube in step 2, with a total volume of 20μl (operate on ice, the reaction system can be expanded as needed):
|
The reaction solution of step (2) |
10μl |
|
5x SuperFast RT Mix |
4μl |
|
DEPC-ddH2O |
6μl |
4. Gently mix the reaction solution, centrifuge briefly to concentrate the solution at the bottom of the tube; incubate at 50°C for 5min; heat at 85℃ for 5s to inactivate the enzyme, and place on ice for subsequent experiments or freeze.
Notes:
1. Please pay attention to avoid RNase contamination during the experiment.
2. When the system after the reverse transcription reaction is completed is used as a template for real-time PCR, it is recommended to add it directly after dilution, and the added volume should not exceed 20% of the PCR reaction system. The heat-resistant DNA polymerase and thermal cycle conditions used for PCR amplification need to be selected according to the size of the target fragment, GC content, primer characteristics, and fidelity requirements.
3. Long-term storage may cause precipitation of the product. Please completely dissolve and mix before use to prevent uneven salt ion concentration from affecting the experimental results.
4. The quality of the RNA template has an important impact on the efficiency of cDNA synthesis. Please choose a reliable RNA extraction/purification method. It is recommended to use TRIzol Reagent (Catalog Number: Such as GK20008 or GK20009) to prepare RNA.
| Components | 50 rxns | 100 rxns |
| 5x gDNA Remover Mix | 100 μL | 200 μL |
| 5x SuperFast RT Mix | 200 μL | 400 μL |
| DEPC-ddH2O | 0.75 mL | 1.5 mL |
| Applications |
A simple and fast reverse transcription kit.
It only takes 8-10 minutes to complete the removal of gDNA and synthesis of cDNA. The obtained cDNA can be directly used as a template for subsequent operations such as Realtime PCR. |
| Shipping | Ship with blue ice. |
| Storage Conditions | Stored at -20°C, and is stable for up to 18 months. Avoid repetitive freeze-thaw cycles. |
| Usage | For research use only! Not for use on humans. |
Average Rating: 5 (Based on Reviews and 30 reference(s) in Google Scholar.)
GLPBIO products are for RESEARCH USE ONLY. Please make sure your review or question is research based.
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