• Designed for cell-free DNA: newly formulated repair and ligation reagents with optimized protocols
• High performance NGS libraries: high diversity with broad and reproducible GC coverage
• Variable sample input: <1 ng to 30 ng of cell-free DNA
• Fast and simple workflow: 3 steps in a single tube or well in 2 hours with no purification or sample transfer steps
• Research applications: libraries from samples where results require high sensitivity including liquid biopsies, ctDNA, and targeted sequencing
• Automation-friendly: ApolloTM Library Prep System(Takara Bio USA) Beckman^® FXP Workstation

제품설명

보관

내용

Component name	Cap Color	12S Kit 12 Single Indexes	48S Kit 48 Single Indexes	96D Kit 96 Dual Indexes
Code		R400490	R400491	R400492
용량		12회	48회	96회
Preparation Buffer	Red	1 Tube	1 Tube	2 Tubes
Template Preparation Enzyme	Red	1 Tube	1 Tube	2 Tubes
Library Synthesis Buffer	Yellow	1 Tube	1 Tube	2 Tubes
Library Synthesis Enzyme	Yellow	1 Tube	1 Tube	2 Tubes
Library Amplification Buffer	Green	1 Tube	1 Tube	2 Tubes
Library Amplification Enzyme	Green	1 Tube	1 Tube	2 Tubes
Nuclease-Free Water	Clear	1 Tube	1 Tube	1 Tube
Indexing Reagents	Blue	12 Tubes	1 Single Index Plate (48S)	1 Dual Index Plate (96D)

※ ThruPLEX^® Plasma-seq Kit single Index 서열
   * ThruPLEX Plasma-seq 12S Kit (R400490)
   * ThruPLEX Plasma-seq 48S Kit (R400491)
※ ThruPLEX^® Plasma-seq Kit Index Guide 바로 가기


[그림1] ThruPlex^® Plasma-Seq Kit Single-tube Workflow
Starting with 1 to 30 ng of cell-free DNA, ThruPLEX^® Plasma-seq Kit creates indexed libraries in 3 simple steps: end repair, adapter ligation, and high-fidelity library amplification. No purification or sample transfer steps are required. The streamlined workflow is performed in 2 hours in a single tube or well, preventing sample loss and enhancing positive sample identification.


[그림2] ThruPlex^® Plasma-Seq Kit원리
ThruPLEX^® Plasma-seq Kit technology is a 3-step reaction that is optimized for cell-free DNA. Cell-free DNA (1 ng to 30 ng) is first repaired in a highly efficient process. Background is reduced using double-stranded adapters with no single-stranded tails. Blunt-end ligation occurs with high-efficiency. Blocked 5’ ends reduce adapter-adapter ligation. Background is further reduced by destroying unused adapters after ligation. A high-fidelity amplification completes the reaction to generate indexed Illumina libraries.


[그림3] Highest Diversity, Fewest Unmapped Reads from Cell-free DNA
ThruPLEX^® Plasma-seq Kit generated quality libraries with high diversity and a low number of duplicates and unmapped reads. Cell-free DNA was extracted from 3 plasma samples, and libraries were prepared at the amounts indicated as measured by Qubit^®. The amount of mononucleosomal DNA in each sample as measured by the Bioanalyzer^® corresponded to 0.09 ng, 0.62 ng, and 15.44 ng. Pooled libraries were sequenced on an Illumina NextSeq^® 500 as a paired-end run with 17M to 25M reads per library, Duplication rates were calculated after down-sampling the data to 17 M reads per library.


[그림4] OutstandingTarget Enrichment Performance
ThruPLEX^® Plasma-seq Kit libraries were captured at high efficiency and generated data with deep coverage of the kinome for mutation detection. Libraries were prepared from 3 plasma samples at input amounts of 5 ng, 6.5 ng, and 10 ng in triplicate, and targeted sequencing was carried out on an Illumina MiSeq^® using samples enriched with the ClearSeq Human DNA Kinome Panel for SureSelectXT2. On average, 5M reads were generated per library. Selected bases were successfully captured bases that were in or within 250 bp of the baits.


[그림5] Reproducible, Unbiased GC Coverage
ThruPLEX^® Plasma-seq Kit provided the most reproducible and unbiased GC coverage across the human genome. ThruPLEX^® libraries showed minimal variability across 9 individual plasma samples tested. Libraries were prepared from cell-free DNA isolated from 1 ml of plasma samples and sequenced on an Illumina NextSeq^® 500. Four separate plasma samples were used to construct the NEBNext^® Ultra libraries.