HiScript III RT SuperMix for qPCR (+gDNA wiper)

Hiscript III RT SuperMix for PCR (+gDNA wiper) is an upgraded vesion of Hiscript Ⅱ Q RT Supermix for GPCR(+gDNA wiper), induding Hiscript Ⅲ Reverse Transcriptase, a new generation of reverse transcriptase with optimized Buffer. This kit further improves the efficiency of DNA synthesis, and is suitable for two-step qRT-PCR detection.

The 4 x DNA wiper Mix in the kit completely removes residual genomic DNA from the RNA template ensuing more reliable quantive results. It simplifies PCR primer design without the need to design primers across introns 5 x Hiscript Ⅲ qRT SuperMix contains all components requried for the reverse transcription reaction and it can be started rapidly after the addition of template RNA and Rnase-free ddH₂O, and the gDNA wiper is terminated to ensure the integrity of the cDNA. It is compatible with dye-based and probe-based qPCR, enabling high-performance gene expression analysis.

Product advantages

• Easy and quick operation: the 5 × HiScript^® III qRT SuperMix contains all the components required for reverse transcription. The reverse transcription reaction can be completed in 20 minutes by adding template RNA
• Broad template compatibility: Compatibility with different species of RNA templates with poor integrity
• Strong tolerance of impurities: for reverse transcription common impurities such as ethanol, isopropyl alcohol, water balance phenol, guanidine isothiocyanate, humic acid, etc., with strong tolerance
• Superior reverse transcription Efficiency: Compared with common commercially available reverse transcription products, the HiScript® III RT SuperMix for qPCR (+gDNA WIper) reverses the smallest quantitative CT value of cDNA and has excellent reverse transcription efficiency

Applications

This product is suitable for reverse transcription of animal, plant and microbial RNA, and the products are compatible with dye-based and probe-based qPCR.

Self-prepared materials

• RNase-free centrifuge tube (1.5 ml), RNase-free PCR tube (0.2 ml), RNase-free tips
• Pipette, PCR instrument, ice or ice box

RNA

High quality RNAis essential for obtaining high quality cDNA. Please verify the RNA integrity by electrophoresis before the experiment.

qPCR Reagent Selection Guide:

The 1st strand cDNA product can be used as the template for gPCR directly. For PCR, it is recommended that the volume of the template cDNA product should not exceed 1/10 of the total volume of qPCR reaction. AceQ Universal U' Probe Master Mix V2 (Vazyme #Q513) or ChamQ Universal SYBR qPCR Master Mix (Vazyme #Q711)can be selected as the qPCR reagent.

Notes

The 4× gDNA wiper Mix,5×HiScriptI qRT SuperMix and 5×No RT Control Mix contain high concentration of glycerol. Please centrifuge briefly and pipette up and down to mix thoroughly before use.

It is recommended to add no more than 1 μg of total RNA to the 20 μl reverse transcription reaction system. If target genes with low expression level, the amount of total RNA can be up to 5 μg. Otherwise, the amount of RNA added is too high, which may willexceed the linear range of subsequent qPCR.

The cDNA products are only suitable for qPCR reactions and not suitable for long-fragment PCR amplification of downstream experi-ments such as cloning. If necessary, HiScript Il 1st Strand cDNA Synthesis Kit(+ gDNA wiper)(Vazyme #R312) is recommended.

Reverse transcription can be performed directly with 5× HiScript I qRT SuperMix without the genome removal step, but do not use 4× gDNA wiper Mix with reagents without genome removal module,as the Mix in the kits without genome removal module does not contain the components to terminate the gDNA wiper reaction, which may affect subsequent gPCR results.