Title: Does the ratio of different RNA & DNA mixtures affect the efficiency of enzymatic digestion?

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.

Title: Does NadPrep RNA & DNA Library Co-Preparation Module support RNA & DNA co-capture?

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.

Title: Is there a difference in performance between the NadPrep RNA & DNA Library Co-Preparation Module and the NadPrep EZ DNA Library Preparation Module v2 when preparing libraries from DNA samples?

Figure 1. Library yield and capture performance of DNA samples using different library preparation protocols. A. Library Yield; B. Mappability.

Note: The sample originated from 50 ng of Human Genomic DNA Standard (Promega, G1521). Sequencing mode was Illumina NovaSeq 6000 platform, PE150.

Title: Is there any difference in the capture performance of libraries prepared from RNA samples using the NadPrep RNA & DNA Library Co-Preparation Module and NadPrep Total RNA-To-DNA Module conjunction NadPrep DNA Library Preparation Module?

Figure 2. Correlation of ERCC expression profiles in captured libraries from RNA samples processed through different library preparation protocols. A. NadPrep RNA & DNA Library Co-Preparation Module; B. NadPrep Total RNA-To-DNA Module conjunction NadPrep DNA Library Preparation Module.

Note: The samples originated from K562 cell line RNA with an initial input of 50 ng; the pre-library input amount was 200 ng.

Title: Is the NadPrep rRNA Fast Blocking Comprehensive Solution suitable for severely degraded FFPE samples? 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%).

Title: Is the NadPrep rRNA Fast Blocking Comprehensive Solution compatible with other third-party RNA library preparation kits? 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.

Title: Why is the dsDNA yield slightly higher when prepared using the NadPrep rRNA Fast Blocking Comprehensive Solution? 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

Title: Can the NadPrep EZ DNA Library Preparation Kit v2 be used after being placed at room temperature overnight?

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.

Title: How many times can the NadPrep EZ DNA Library Preparation Kit v2 be frozen and thawed?

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.

Title: Is it appropriate to use input DNA that is higher or lower than recommended in the user manual?

In order to ensure the data quality, it is best to prepare the library within the scope recommended in the user manual. If there are special experimental needs, it can also be adjusted appropriately. The libraries prepared fromdifferent input DNA areshown as follows:

Experimental conditions: Human Genomic DNA Male (Promega, catalog # G1471) was fragmented with different input DNA, with 25 min of the enzymatic digestion time. NadPrep Universal Stubby Adapter (UDI) Module was used together for library preparation with single-sided selection mode. The library yield and size meet the requirements of sequencing.