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Which sample types are the NadPrep EZ RNA Library Preparation Kit applicable for?
Title: Which sample types are the NadPrep EZ RNA Library Preparation Kit applicable for? NadPrep EZ RNA Library Preparation Kit is widely suitable for the libraries prepared from total RNA with different grades, poly(A)-enriched mRNA, and rRNA-depleted RNA. It should be noted that the mRNA content in total RNA from different sources varies greatly. If the initial input amount of total RNA is too low, sufficient mRNA may not be obtained to prepare a high-quality library.
What’s the difference between non-stranded and stranded libraries from NadPrep EZ RNA Library Preparation Kit?
Title: What’s the difference between non-stranded and stranded libraries from NadPrep EZ RNA Library Preparation Kit? RNA-Seq library preparation generally falls into two types: non-stranded (general) and stranded (strand-specific). The core difference is whether the library retains transcript directional information. Among them, stranded libraries retain the transcript directional information during the preparing process, and sequencing and downstream analysis can determine whether the transcripts originate from the sense or antisense strands of DNA. However, non-stranded libraries lose directional information during cDNA synthesis, and sequencing data cannot directly indicate transcript directions. Compared with general RNA-Seq, strand-specific RNA-Seq provides more accurate transcript quantification, clearer gene structures, identification of antisense transcripts, and discovery of novel transcripts. Therefore, stranded RNA-Seq is crucial for studies of gene structure and expression regulation. To obtain transcript direction information, stranded libraries should be preferred.
Can RNA samples be directly used for library preparation with this kit?
Title: Can RNA samples be directly used for library preparation with this kit?

No. This kit is compatible only with gDNA or cDNA as initial samples. For RNA samples, reverse transcription to generate cDNA is required prior to library preparation.

What is the recommended input range? How to handle inputs exceeding this range?
Title: What is the recommended input range? How to handle inputs exceeding this range?

This kit supports 50-2,000 ng of gDNA or cDNA. For input amount exceeds 2,000 ng, split the sample into multiple reactions to maintain amplification efficiency.

What is the insert size of this kit? How to select sequencing read length?
Title: What is the insert size of this kit? How to select sequencing read length?

The main peak of PCR products by using this kit is ~270 bp. PE150 sequencing is recommended for high-quality coverage.

What is the recommended sequencing data volume for IGTR immune repertoire analysis?
Title: What is the recommended sequencing data volume for IGTR immune repertoire analysis?

A minimum of 0.3 Gb is recommended to detect clones at 0.01% abundance with an input of 200 ng. Increase data volume to enhance sensitivity for low-frequency clones.

Is this kit suitable for minimal residual disease (MRD) monitoring?
Title: Is this kit suitable for minimal residual disease (MRD) monitoring?

Yes. This kit includes IG Primer Mix and TR Primer Mix with gene-specific primers provided in separate tubes, allowing flexible combination in a single amplification reaction. With its high sensitivity, the kit meets the requirements for MRD monitoring technology development and clinical applications, making it ideal for low-frequency variant detection scenarios.

How to deal with low yield of RNA & DNA co-prepared library?
Title: How to deal with low yield of RNA & DNA co-prepared library?

√ 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.

√ If the nucleic acid sample contains excessive amounts of chelating agents, guanidine salts, phenol, proteins, ethanol, or other impurities, it can impact the activity of RNA reverse transcriptase and the efficiency of DNA enzymatic digestion. It is recommended refer to purify the nucleic acid samples using 2X NadPrep SP Beads, replace the elution reagent with Nuclease Free Water, and then proceed with library preparation.

√ For samples of lower quality, consider increasing the number of PCR amplification cycles appropriately.


How to deal with the issue of excessively large or double peaks in RNA & DNA Co-prepared library fragments?
Title: How to deal with the issue of excessively large or double peaks in RNA & DNA Co-prepared library fragments?

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.

Can NadPrep RNA & DNA Library Co-Preparation Module be compatible with EDTA-eluted samples?
Title: Can NadPrep RNA & DNA Library Co-Preparation Module be compatible with EDTA-eluted samples?

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.

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