<i>Evo M-MLV</i> 反转录预混型试剂盒（ 含去除 gDNA 试剂, 用于 qPCR ）Ver.2

Evo M-MLV Reverse Transcription Premix Kit (with gDNA removal reagent for qPCR) Ver.2

Product No.:AG11728

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¥1,500

Product Description

This product utilizes M-MLV Reverse transcription kit for reverse transcriptase. cDNA products are ready for qPCR. 5X Evo M-MLV RT Reaction Mix Ver.2 contains all the components required for the reverse transcription reaction and is optimized for Oligo dT (18T) Primer, Random 6 mers Primer dosage, making the reverse transcription product compatible with both dye-embedded and probe-based qPCR assays for efficient gene expression analysis.
In downstream quantitative PCR experiments, the genomic DNA (gDNA) mixed in Total RNA can be used directly as the template for amplification, which may cause false positive results. The gDNA Clean Reaction Mix Ver.2 effectively removes the residual genomic DNA to ensure the accuracy of quantitative results. Meanwhile, the product is equipped with NRT Control Reaction Mix, which can be used to prepare a negative control reaction without reverse transcriptase.
Evo M-MLV The Reverse Transcription Premix Kit (with gDNA removal reagents for qPCR) Ver.2 is a new version of the original Evo M-MLV Reverse transcription pre-mixed kit (with gDNA removal reagent for qPCR) is optimized and upgraded on the basis of different operation methods, customers can choose according to their needs:

1) Method 1: Two-step method (remove gDNA first, then reverse transcription): This method is recommended if there is too much gDNA residue in the extracted RNA or if there is a higher demand for gDNA removal.
2) Method 2: All in one (remove gDNA and reverse transcription reaction in one tube at the same time): When the quality of RNA is high (less gDNA residue), or the concentration of RNA in the reaction is high, and the copy number of the target gene is high, this method can be chosen, which is simple and fast, and it can simplify the experimental steps and shorten the length of the experiment.

Product Advantages

① Contains gDNA Clean Reaction Mix Ver.2, a genomic DNA removal reagent, which can quickly remove genomic DNA mixed in RNA templates.
② The kit will remove the genome and reverse transcription system separately prepared into a premix, easy to use, can reduce the loss of reagents and experimental errors.
③ Applicable to different operation methods, customers can choose according to their needs: either choose the two-step method, first remove the gDNA, and then reverse transcription; or choose All in one, complete the removal of the genome and reverse transcription reaction in one tube at the same time, the operation is convenient and fast.
④ The cDNA obtained from this product is suitable for both SYBR and probe qPCR analysis.

Product Composition
individual parts making up a compound norm
gDNA Clean Reaction Mix Ver.2 *1 240 μl
5X Evo M-MLV RT Reaction Mix Ver.2*2 400 μl
NRT Control Reaction Mix*3 80 μl
RNase free water 1 ml X 2 pcs

*1: gDNA Clean Reaction Mix Ver.2 can be used with 5X. Evo M-MLV Use with RT Reaction Mix Ver.2 or NRT Control Reaction Mix. gDNA Clean Reaction Mix Ver.2 will be left over if not used with NRT Control Reaction Mix.
*2: The solution contains Evo M-MLV RTase, RNase Inhibitor, dNTPs, Oligo dT (18T) Primer and Random 6 mers Primer.
*3: The solution does not contain Evo M-MLV RTase, and the remaining fractions with 5X Evo M-MLV RT Reaction Mix Ver.2 is identical.

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

Add 200 ng of Human genomic DNA to 293T Cell Total RNA (200 ng, 20 ng, 2 ng, 200 pg, 20 pg, 2 pg, 0 pg, respectively) and use the product to remove the genomic DNA, followed by reverse transcription.(Methodology I: two-step system 1)The cDNA was used as a template, and 2 μl of the cDNA stock solution was extracted and analyzed using our product SYBR Green. Pro Taq HS Premix qPCR kit (Code No. AG11701) was used for qPCR amplification detection of Human Actin Genes.
Quantitative instrument used: ABI QuantStudioTM 5 Real-Time PCR Systems. the results are shown below:

The results are shown above:
1、Working curve R2= 0.9987 and the amplification efficiency is 101.41 TP3T.
In the samples without RNA but with gDNA, the Ct value was not detected within 35 cycles.
3. The product has high reversal efficiency and can be used for accurate quantification over a wide template range. The cDNA amplification curves obtained by reverse transcription of 200 ng ~ 2 pg Total RNA show a good linear relationship.
4、Single peak type in melting curve, good amplification specificity.

Example 2

Mouse Liver Total RNA was reverse transcribed using this product (Method 2: All in oneThe starting amount of Total RNA was 1 μg, 100 ng, 10 ng, 1 ng, 100 pg, 10 pg, 1 pg. 2 μl of cDNA stock solution was taken as a template for the cDNA of reverse transcription product, and the cDNA was extracted from the cDNA stock solution using our product SYBR Green. Pro Taq HS Premix qPCR kit (Code No. AG11701) was used for qPCR amplification to detect Mouse's GAPDH Genes.
Quantitative instrument used: ABI QuantStudioTM 5 Real-Time PCR Systems. the results are shown below:

The results are shown above:
① Working curve R2= 0.9959 and the amplification efficiency was 106.71 TP3T.
② 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 1 μg ~ 1 pg Total RNA show good linearity.
③ The melting curve has a single peak type and good amplification specificity.

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