Store at -20°C,Avoid repeated freezing and thawing
Shipped In
Ice chest + Ice pads
Product Description
Product content:
N670210
Component
16×10 μL
Storage
N670210A
Ion P1 Adapter (10 μM)
160 μL
-20℃. Avoid freeze/thaw cycle.
N670210B
Ion Barcode Ax (10 μM)
16×10 μL
-20℃. Avoid freeze/thaw cycle.
Product Introduction:
NGS Barcode Adapter for Ion Torrent (1-16) is a specialized kit designed for library construction on Ion Torrent high-throughput sequencing platforms. It can be used to construct libraries suitable for Ion PGM ™ System, Ion Proton System, Ion S5 System, and Ion GeneStudio ™ Multi sample targeted sequencing DNA library using Ion Torrent high-throughput sequencing platforms such as S5 Series System. This kit contains 16 different barcode adapters, including P1 Adaptor and Barcode A1 to Barcode A16, used in the library construction of the Ion Torrent sequencing platform. It is compatible with the Kangwei Century second-generation sequencing rapid DNA library construction kit and the AmpliSeq second-generation sequencing DNA library construction kit and can be used to construct 16 index labeled DNA libraries for sequencing analysis. All reagents provided in the kit have undergone strict quality control and functional validation, ensuring the reliability of sample identification, stability and repeatability of library construction to the greatest extent possible.
Notice:
1. The adapter is a double chain structure, please do not place it above room temperature to avoid chain breakage and affect use.
2. Before opening the barcode adapter, please centrifuge briefly to collect the liquid at the bottom of the tube to avoid cross contamination between different barcode adapters.
Operation steps The packaging quantity of each Barcode Adapter in this kit is sufficient for library construction of ≤ 14 pieces of 100 ng Input DNA or 1 piece of 1 µ g Input DNA. The amount of adapter used can be adjusted according to the initial amount of DNA in the sample to achieve optimal usage effect. The following operation example is performed using the Kangwei Century second-generation sequencing rapid DNA library kit:
1. Add the following reagents sequentially into a new 0.2 ml PCR tube and mix thoroughly.
Component
Volume
Volume
50-100-ng DNA Input
1-µg DNA Input
End Repaired DNA Fragment
60 μl
60 μl
Ligation and Nick Repair Buffer
10 μl
10 μl
Ion P1 Adapter
7 μl(1 μM )
7 μl(10 μM )
Ion Barcode Ax
7 μl(1 μM )
7 μl(10 μM )
Nuclease-free Water
12 μl
10 μl
DNA Ligase
2 μl
4 μl
Bst DNA Polymerase
2 μl
2 μl
Total volume
100 μl
100 μl
Attention: It is recommended that the molar ratio of the amount of Adaptor added to the DNA fragment be 10:1-20:1. Please refer to the instructions for using the Kangwei Century second-generation sequencing rapid DNA library kit for specific concentrations and amounts of Adaptor. When the DNA amount is 10-100 ng, the recommended concentration of Adaptor is 1 μ M (less than 260 bp) or 0.5 μ M (300-400 bp); When the DNA content is 1 µ g, Adaptor recommends using a concentration of 10 μ M (less than 260 bp) or 5 μ M (300-400 bp). 2. Place the PCR tube in the PCR instrument, set the hot cap to 80 ℃, and run the following program:
Stage
Temperature
Time
Hold
25℃
15 minutes
Hold
65℃
5 minutes
Hold
4℃
Hold
Attention: Please proceed to the next step as soon as possible after connecting the connector to purify the product of the connector connection.
Sequence information Ion P1 Adapter：5’-CCACTACGCCTCCGCTTTCCTCTCTATGGGCAGTCGGTGAT-3’ Ion Barcode Ax：5’-CCATCTCATCCCTGCGTGTCTCCGACTCAGXXXXXXXXXXGAT-3’ *The underlined sequence X is the barcode sequence used during sequencing. The sequence of barcodes corresponding to each Barcode Adapter is as follows:
