AccuNext  Chain-Specific Single Cell/Micro RNA Construction Kit II (Illumina)

This product is a strand-specific RNA sequencing library construction kit designed for illumina's high-throughput sequencing platform. It can be used to construct RNA libraries from 1~100 cells or 10 pg~10 ng Total RNA samples, and contains all the components required for RNA fragmentation, reverse transcription (RT) and PCR library amplification. The product fragments RNA by heating, then reverse transcribes it with a 3′-N6 Primer with a universal junction sequence, which utilizes the terminal transferase activity of the reverse transcriptase enzyme to add a specific junction to the end of the cDNA; PCR amplification is then performed with primers with illumina sequencing junctions [for example.AccuNext CDI Junction Primers (Illumina, for RNA libraries) (Code No. AG12505, AG12506, AG12507)], purification can be completed to obtain sequencing libraries for illumina sequencing platform.
This product is easy to operate and can obtain RNA sequencing libraries without the traditional tedious steps such as end repair and splice connection; the optimized reaction system improves the efficiency of library conversion, and the data obtained from sequencing is of high quality with complete coverage; at the same time, the template conversion reaction retains the directionality of the RNA strand, and RNA strand-specific sequencing data can be obtained.
Please use the reagents provided in this product for RNA fragmentation, reverse transcription and PCR library amplification experiments. It is not recommended to change the dosage or concentration of any of the reaction components or to replace the components in this product with other equivalent products in order to avoid obtaining bad results. If substitution is necessary, please verify first.
Since rRNA accounts for nearly 90% of the total RNA, it is recommended to remove the rRNA before constructing the RNA library in order to obtain more effective data volume during sequencing and to reduce the sequencing cost.
The rRNA removal module is not included in this product, you can choose the appropriate rRNA removal reagent according to your needs. If you do not need to remove rRNA or if you have already removed rRNA, you can directly follow the instructions of this product; if you need to remove rRNA, you can purchase our product AccuNext Chain-Specific Single-Cell/Micro RNA Construction Kit (Illumina) (Code No. AG12526, AG12527, Note: This product contains a module for removing rRNA from human samples). (Code No. AG12526, AG12527, Note: This product contains a module for removing rRNA from human samples).

1. For illumina sequencing platform: integrating Index and illumina connectors, constructed RNA libraries can be directly sequenced by NGS.
2. High sensitivity: RNA library construction from single cells or 10 pg Total RNA.
3. Wide range of applications: suitable for RNA samples with RIN values from 2 to 10, and at the same time, compatible with degraded samples (e.g., FFPE samples).
4. Simple operation: RNA sequencing libraries can be obtained without the traditional tedious steps such as end repair and junction connection.
5. Accurately recognizes RNA strand specificity: Read 1 reads the antisense strand sequence of the original RNA, and Read 2 reads the positive-sense strand sequence of the original RNA.

Component (Package 2-1, stored at -80°C)	AG12543	AG12544	AG12545
5′- Adapter Primer Mix	54 μl	216 μl	216 μl X 2 pcs
Control Total RNA (1 μg/μl)*1	5 μl	5 μl	5 μl
Components (Package 2-2, stored at -20°C)
10X Cell Lysis Buffer	228 μl	920 μl	920 μl X 2 pcs
RNase Inhibitor ( 40 U/μl )	18 μl	72 μl	144 μl
5X First-Strand Buffer	48 μl	192 μl	384 μl
3′-N6 Primer	12 μl	48 μl	96 μl
AccuNext Reverse Transcriptase (100 U/μl)	24 μl	96 μl	192 μl
AccuNext DNA Polymerase (1 U/μl )	36 μl	144 μl	288 μl
2X AccuNext PCR Buffer	900 μl	900 μl X 4 pcs	1.2 ml X 6 pcs
AccuNext PCR Primer Mix	24 μl	96 μl	192 μl
Nuclease free water	1 ml	1 ml X 3 pcs	1 ml X 6 pcs

*1: Control Total RNA is 293T Cell Total RNA.
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.

Storage temperature: Package 2-1 -80℃ Storage Package 2-2 -20℃ Storage
Transportation Temperature: Package 2-1 Dry Ice Transportation Package 2-2 Dry Ice Transportation or -20℃ Ice Pack Transportation