<i>Evo M-MLV</i> 反转录试剂盒（含去除 gDNA 试剂, 用于 qPCR）

Evo M-MLV Reverse transcription kit (contains gDNA removal reagent, for qPCR)

Product Code: AG11705

- +

¥1,400

Product Description

This product is a special reagent that removes genomic DNA mixed in cDNA for Real-Time RT-PCR reaction. The genomic DNA will affect the quantitative analysis of cDNA. gDNA Clean Reagent, which has strong DNA decomposition activity, is used in this product to remove the genomic DNA in 2 min at 42 ℃. cDNA can be synthesized from the processed samples by subsequent reverse transcription reaction and then analyzed by quantitative PCR. The cDNA synthesized by this product is suitable for chimeric and probe-based qPCR analysis.

Product Advantages

The gDNA Clean Reagent is a genomic DNA removal reagent that removes genomic DNA from RNA templates quickly.
② The OligodT (18T) Primer (50 μM) and Random 6 mers Primer (400 μM) are included in the kit, so that different primers can be selected for reverse transcription according to the actual situation, and the concentration of the primers can also be adjusted according to the needs.
③ The cDNA obtained by inversion of this product can be used for both SYBR and probe qPCR analysis.

Product Composition
individual parts making up a compound norm
gDNA Clean Reagent 100 μl
5X gDNA Clean Buffer 200 μl
Evo M-MLV RTase Enzyme Mix *1 100 μl
5X RTase Reaction Buffer Mix I*2 400 μl
Oligo dT (18T) Primer (50 μM) 100 μl
Random 6 mers Primer (400 μM) 100 μl
RNase free water 1 ml x 2 pc

*1: Contains RNase Inhibitor.
*2: Contains dNTP.

 

Preservation and transportation
Storage temperature: -20℃
Transportation temperature: dry ice or -20℃ ice bag
Example 1

Add 200 ng of mouse genomic DNA to mouse liver total RNA (2 μg, 200 ng, 20 ng, 2 ng, 200 pg, 20 pg, 2 pg, 0 pg, respectively), and then use this kit to perform the genomic DNA removal reaction, and then carry out the reverse transcription experiments after removing the genomic DNA, and then take 2 μl of cDNA as a template. The cDNA was used as a template, and 2 μl of the cDNA stock solution was taken and analyzed by SYBR Green Premix. Pro Taq HS qPCR Kit (Code. AG11701) for qPCR amplification to detect mouse GAPDH Genes.

The results are shown above:
① Working curve R2= 0.996, amplification efficiency 103.11 TP3T.
② No RNA but gDNA samples were added, and the Ct value was not detected within 35 cycles.
③ This product has a high reversal efficiency and can be accurately quantified over a wide template range. The cDNA amplification curves obtained by reverse transcription of 2 μg ~ 2 pg Total RNA show good linearity.
④ The melting curve has a single peak type and good amplification specificity.

Example 2

Add 200 ng of human genomic DNA to 293T cell Total RNA (2 μg, 200 ng, 20 ng, 2 ng, 200 pg, 20 pg, 2 pg, 0 pg, respectively), and then perform the genomic DNA removal reaction using this kit. Using this cDNA as a template, take 2 μl of cDNA stock solution, and use SYBR Green Premix to remove the cDNA. Pro Taq Amplification by HS qPCR Kit (Code. AG11701) GAPDH Genes.

The results are shown above:
①Working curve R2= 0.995, amplification efficiency 103.0%.
In the samples without RNA but with gDNA, the Ct value was not detected within 35 cycles.
③This product has a high reversal efficiency and can be accurately quantified over a wide template range, and the cDNA amplification curve obtained by reverse transcription of 2 μg ~ 2 pg Total RNA shows good linearity.
④ The melting curve has a single peak type and good amplification specificity.

Example 3

Mouse liver Total RNA was used as the template for genomic DNA removal reaction and reverse transcription experiments using this kit. 2 μg~2 pg of RNA starting template was used. 2 μl of the cDNA stock solution was taken, and the cDNA was extracted from the cDNA with the Pro Taq The HS Premix Probe qPCR Kit (Code. AG11704) uses probe-based qPCR to amplify mouse GAPDH Genes.

The results are shown above:
①Working curve R2= 0.997 with an amplification efficiency of 104.01 TP3T;
②This product has a high reversal efficiency and can be accurately quantified over a wide template range. The cDNA amplification curves obtained by reverse transcription of 2 μg ~ 2 pg Total RNA show good linearity.

Related products