Libraries prepared with this module can be used in conjunction with NEX-t Panel v1.0 and NadPrep ES Hybrid Capture Reagents. This significantly shortens the time required for pathogen-targeted sequencing and simplifies experimental procedures.

When co-prepare RNA & DNA libraries from mixed nucleic acid samples, a higher proportion of DNA samples results in larger fragments with enzymatic digestion. Adjustments can be made according to the expected insert fragment lengths and recommended time of enzymatic fragmentation in the manual.

This module can tolerate nucleic acid samples containing 6 μL of EDTA (1 mM), resulting in a final concentration of 0.3 mM in the cDNA first-strand synthesis system. At this concentration, RNA reverse transcription inhibition is minimal, and the efficiency of enzymatic digestion is unaffected. Before library preparation, confirm the sample solvent. If it exceeds the recommended EDTA threshold, it is necessary to purify nucleic acid samples using 2X NadPrep SP Beads, replacing the elution reagent with Nuclease Free Water before proceeding with library preparation.

If the nucleic acid samples contain high concentrations of EDTA or have an excessively high pH, it can affect the efficiency of DNA digestion. It is recommended to purify nucleic acid samples using 2X NadPrep SP Beads, replacing the elution reagent with Nuclease Free Water before proceeding with library preparation.

√ If the sample is a co-extracted total nucleic acid, quantification of RNA and DNA should be performed separately using Qubit, and library amplification should be carried out according to the recommended cycles in the manual. 

√ During the experiment, RNase-free consumables must be used, and the operating environment should be free of RNase contamination, as this can affect the efficiency of RNA reverse transcription.

A: This solution is suitable for RNA samples from a wide range of cell and tissue sources with varying degradation levels. However, it is not recommended for severely degraded FFPE samples (DV 200 < 20%).

A: Yes, it is. This solution can be integrated with RNA library preparation kits for the preparation of total RNA sequencing (RNA-Seq) libraries. If you choose a third-party RNA library preparation kit, we recommend using NEBNext® Ultra™ II RNA Library Prep Kit for Illumina® (NEB, Cat # E7770L) or the KAPA RNA HyperPrep Kit (Roche, Cat # KK8541). For the use of RNA library preparation kits from other brands, please consult technical support from Nanodigmbio with support@njnad.com. 

A: This solution contains a large amount of rRNA ssDNA blocking reagent. When used in conjunction with the NadPrep Total RNA-To-DNA Module, there may be a slight residual of ssDNA in the purified dsDNA product. When quantifying dsDNA using a nucleic acid fluorescence quantifier, this residual ssDNA can influence the results, leading to a slightly higher yield. However, the remaining rRNA ssDNA blocking reagent does not affect subsequent experiments. We recommend appropriate adapter dilution factors and PCR amplification cycles for different different types of RNA and different input amounts. 

The kit can be used after being placed on the table at room temperature overnight without UV light irradiation. NadPrep EZ DNA Library Preparation Kit v2 hasa stable performance even at room temperature 12 h.

NadPrep EZ DNA Library Preparation Kit v2 is recommended to be frozen and thawed within 15 times. In order to ensure the data quality, it is recommended to use it in sub packaging. Please mix well before sub packaging.