The Low EDTA TE buffer and nuclease-free water can be stored at room temperature. All other components should be stored at -20°C. The shelf life of all reagents is one year when stored properly.
|Tube name||8 x RXN||24 x RXN||96 x RXN|
|End preparation||End Prep buffer|
End Prep enzymes
|Adaptor ligation||Ligase MM|
Low EDTA TE
|Amplification||2x PCR master mix|
01. To enable multiplexing, please use singleplex or multiplex adapter for illumina.
02. Please you use high quality DNA as possible as you can. Heavily nicked or damaged DNA may cause library preparation failure. You can use 260 nm/280 nm ratios (1.8~2.0) to check DNA purity. You also can use fluorescent method to perform double-stranded DNA quantification. Contaminating RNA, nucleotides and single-stranded DNA may affect the amount of usable DNA in a sample preparation.
03. Please mix by gently pipetting the reaction mix up and down.
The Rapid DNA library preparation kit has been optimized and validated using 1 ng ~ 1 μg of fragmented DNA.
After adaptor ligation, clean up step is designed to remove low molecular weight materials. Size selection step is also designed for users who want to perform size selection for their libraries. If your input DNA is less than 50 ng, size selection is not recommended.
The kit also can be used to perform PCR-free DNA library preparation, in which adaptors contain the complete sequence required for hybridization of the templates to the flowcell surface, and for annealing of sequencing primers. You also can use customized or other commercial PCR-free Y-shape adaptors.
In the amplification step, you also can use yourself-selected DNA polymerase, and PCR primers are included in this kit.
Primer 1: 5’-AATGATACGGCGACCACCGAG
Primer 2: 5’-CAAGCAGAAGACGGCATACGAG
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