AccuNext Small RNA Library Preparation Kit (Illumina)

Item No.: AG12511 / AG12512

- +

Price range: ¥6,800 through ¥20,800

Product Description
This product is a Small RNA sequencing library preparation kit designed for illumina's high-throughput sequencing platform, which can prepare Small RNA sequencing libraries starting from 1 ng~2 μg of Total RNA (containing Small RNA) or 1 ng~1 μg of enriched Small RNA for analyzing a wide range of small RNAs (including miRNA. piRNA, snoRNA and snRNA, etc.), piRNA, snoRNA and snRNA, etc.). Poly (A) Polymerase is used to add a Poly (A) tail to the 3’ end of the RNA, and Oligo (dT) primers with specific junctions are used for reverse transcription, which utilizes the terminal transferase activity of the reverse transcriptase to add specific junctions to the end of the cDNA. PCR amplification with primers for illumina sequencing junctions [e.g.AccuNext CDI Junction Primers (Illumina, for RNA libraries) (Code No. AG12505, AG12506, AG12507)] were used to obtain small RNA libraries of 150 bp ~ 350 bp for illumina sequencing platform by magnetic bead sorting.
The product is easy to operate, without the need to add illumina sequencing junctions and index sequences to obtain Small RNA libraries without the need to carry out tedious steps such as junction connection; the optimized reaction system improves the efficiency of library transformation, is compatible with different template amounts, and obtains stable sequencing results.
This product contains all the components required for RNA 3’ end plus Poly (A) tail, reverse transcription (RT) and PCR amplification, the reaction system has been carefully optimized, all the reagents should be used in the experiments, it is not recommended to change the dosage and concentration of any reaction components or replace them with other equivalent products, in order to avoid obtaining bad results. If replacement is needed, please verify it first.
Product Advantages
1. For Illumina Sequencing Platform: Integrates Index and Illumina connectors to build Small RNA libraries that can be directly sequenced on the Illumina High-Throughput Sequencing Platform.
2. Wide range of templates: able to analyze a variety of Small RNAs including miRNA, piRNA, snoRNA and snRNA from 1 ng~2 μg Total RNA (containing Small RNA) or 1 ng~1 μg enriched Small RNA.
3. Low preference, easy to use, no need for tedious steps such as junctions to add illumina junctions and index sequences to obtain Small RNA sequencing libraries, avoiding the preferences associated with junction ligation.
Product Composition

Package 2-1 components are as follows (stored at -80°C):

individual parts making up a compound AG12511 (12 rxns) AG12512 (48 rxns)
Control micRNA (2.5 ng / μl) 5 μl 5 μl
smRNA Adapter Mix Solution 78 μl 312 μl

Package 2-2 is grouped as follows (stored at -20°C):

individual parts making up a compound AG12511 (12 rxns) AG12512 (48 rxns)
Poly(A) Polymerase (0.5 U/μl) 12 μl 48 μl
4X Poly(A) Reaction Buffer 30 μl 120 μl
RNase Inhibitor ( 40 U/μl ) 9 μl 36 μl
3’smRNA dT Primer 12 μl 48 μl
AccuNext Reverse Transcriptase ( 100 U/μl ) 24 μl 96 μl
2X AccuNext PCR Mix I 600 μl 1.2 ml X 2 pcs
Nuclease free water 1 ml X 2 pcs 1 ml X 5 pcs
Tris Buffer (5 mM) 900 μl 1.2 ml X 3 pcs

Attention:AccuNext CDI Junction Primers (Illumina, for RNA libraries) (Code No. AG12505, AG12506, AG12507) are required for the experiment, but are not configured in this product and need to be purchased separately.AccuNext Information on CDI-joining primers (Illumina, for RNA libraries) is provided in Appendix A and B.

Preservation and transportation
Storage temperature: Package 2-1 -80℃ Storage Package 2-2 -20℃ Storage
Transportation Temperature: Package 2-1 Dry Ice Transportation Package 2-2 -20℃ Ice Pack Transportation or Dry Ice Transportation
Experiment example 1

Mouse liver Total RNA library was constructed by using this product, the amount of RNA template was 100 ng, the number of PCR amplification cycles was 15, and the results of library products after purification and sorting were as follows: Fig. 6-A shows the amplified library products, which were uniformly distributed, with peaks in the range of 150~350 bp. Figure 6-B shows the negative control without template.

 

Example 2

Mouse Liver Total RNA and Mouse Liver Small RNA were used as templates for library construction using this product, and sequenced using the Illumina NovaSeq 6000 system:

 

Example 3

Using 100 ng of murine liver Small RNA and murine liver Total RNA as templates, small RNA libraries were constructed with this product and sequenced using the Illumina NovaSeq 6000 system, and the results showed that the experiments were reproducible (Figure 7-A shows 100 ng of murine liver Small RNA and Figure 7-B shows 100 ng of murine liver Total RNA). The results showed that the experiments were reproducible (Figure 7-A, 100 ng of mouse liver small RNA; Figure 7-B, 100 ng of mouse liver Total RNA).