Products / NGS Lib Prep Kit

Rapid DNA Lib Prep kit

Summary
Catalog No: RK20200
Input DNA amount: 1 ng-1 μg
Size: 8 RXN / 24 RXN / 96 RXN
Sequencer: Illumina

To buy,select size,quantity

Overview

The Rapid DNA library preparation kit is designed for illumina DNA sequencing library construction. The kit 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. The minimal input for preparing PCR-free library is 100 ng of high quality genomic DNA, or 10 ng circulating cell-free DNA (cfDNA). PCR primers are included for users who wish to use self-selected DNA polymerase in the library amplification.
The entire workflow of Rapid DNA library preparation kit is shown as the below.

List of components


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 name8 x RXN24 x RXN96 x RXN
End preparationEnd Prep buffer
End Prep enzymes
100 μl
30 μl
260 μl
78 μl
1000 μl
300 μl
Adaptor ligationLigase MM
Ligase mix
Low EDTA TE
165 μl
30 μl
1000 μl
429 μl
78 μl
2 mL
1650 μl
300 μl
5 mL
Amplification2x PCR master mix
PCR primers
250 μl
15 μl
650 μl
39 μl
2.5 mL
150 μl

Additional Materials required


  • Singleplex or multiplex adaptors for illumina.
  • 80% ethanol (freshly prepared, room temperature).
  • PCR tubes or plates.
  • Magnetic stand.
  • Thermocycler.
  • Agencourt AMPure XP bead (room temperature).
  • Pipettes/multichannel pipettes (2.5, 10, 20, 200, 1000 μl).
  • Minicentrifugator.
  • Vortex.

General Consideration


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.

Protocol


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.

Oligonucleotide Sequences


Adaptor:


Primer 1:  5’-AATGATACGGCGACCACCGAG

Primer 2:  5’-CAAGCAGAAGACGGCATACGAG

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