Description
The Rapid CE DNA Lib Prep kit for Illumina V2 provides a streamlined and efficient method for the construction of DNA libraries for next generation sequencing (NGS) on Illumina® platforms. This kit is an updated version of the original Rapid CE DNA Lib Prep kit (Cat. Nos. RK20212) that has been further optimized to provide improved PCR amplification efficiency. The entire protocol can be completed within 2 hours. Starting from fragmented input DNA (1 - 1000 ng), the kit contains all necessary enzymes and buffers for end-preparation, adapter ligation and library amplification. To simplify the workflow, the fragmented DNA is end-repaired and adenylated during one initial step. The kit can be conveniently used for the construction of a PCR-free library from either 100 ng of high-quality fragmented genomic DNA or 10 ng circulating cell-free DNA (cfDNA). Optional PCR primers are provided in the kit for users who prefer self-selected high-fidelity DNA polymerases for library amplification.
Product Features
- Easy to operate and the entire protocol can be completed in 2.5 hours;
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Applicable to common genomes, cfDNA/ctDNA, FFPE DNA, ChIP DNA, PCR products, cDNA products, etc;
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Wide range of applications for whole genome sequencing, exon and target sequencing, ChIP seq, amplicon sequencing;
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Strict quality control including component pollution test, functional test verification, application scenario test and stability test between product batches.
Workflow Chart
Storage
All components should be stored at -20°C.
The shelf lives of all reagents are one year when stored properly.
Product Components
|
Tube name |
8 RXN |
24 RXN |
96 RXN |
End Preparation |
End Prep Buffer |
80 μL |
240 μL |
960 μL |
End Prep Enzymes |
24 μL |
72 μL |
288 μL |
Adaptor Ligation |
Ligase MM |
132 μL |
396 μL |
1584 μL |
Ligase Mix |
24 μL |
72 μL |
288 μL |
Amplification |
Gloria Nova HS 2X PCR Mix for NGS |
200 μL |
600 μL |
2400 μL |
10X PCR Primers |
40 μL |
120 μL |
480 μL |
Product Data
1. Good species compatibility
GDNA from different sources was used as template, and RK20254 was used for DNA library construction. Except for FFPE, the input amount of DNA was 100 ng, and the library was amplified by 8 cycles. The results showed that for different template types, the yield of the library constructed by RK20254 met the standard of on-machine sequencing.
Using four different FFPE DNA samples as templates, ABRapid CE DNA Lib Prep Kit V2 has a higher yield than similar products.
2. Broad starting input
Different starting amounts of Blood and Rice gDNA were used as templates for DNA library construction using RK20254. The results showed that the libraries constructed by RK20254 had high yield and could be compatible with a wider range of sample starting quantities.
3. Excellent library yield
DNA from different species was used as template, and RK20254 was used for DNA library construction. Except for FFPE, the input amount of DNA was 100 ng, and the library was amplified by 8 cycles. The results showed that the yield of the library constructed by RK20254 was higher than that of its competitors.
4. Excellent sequencing data quality
sample |
product |
Fragment size |
The original Reads |
Proportion of valid data |
GC content |
>Q20 |
>Q30 |
Mapping ratio |
human blood gDNA |
Rapid CE V2 |
186 |
1733332 |
99.31% |
42.05% |
97.56% |
93.08% |
97.76% |
V* |
196 |
1733332 |
99.10% |
41.75% |
97.40% |
92.69% |
97.73% |
Y* |
189 |
1733332 |
98.99% |
42.41% |
97.25% |
92.37% |
97.20% |
Human Blood gDNA was used as template, and RK20254 and competing products were used for library construction and sequencing, respectively. The sequencing results showed that the sequencing quality of RK20254 was consistent compared with its competitors.
5. Excellent accuracy
DNA libraries were constructed and sequenced using RK20254 using DNA templates from different species: human whole blood genome, rice, mouse, E.coli. The results showed that the library constructed by RK20254 was more accurate than its competitors.
6. Excellent capture sequencing quality
sample |
product |
The original Reads |
Proportion of valid data |
GC content |
>Q20 |
>Q30 |
Mapping proportion |
Target proportion |
human blood gDNA |
Rapid CE V2 |
2133332 |
99.32% |
43.69% |
97.20% |
92.40% |
99.79% |
70.23% |
FFPE DNA-liver |
Rapid CE V2 |
2671190 |
99.33% |
49.49% |
97.79% |
93.67% |
99.98% |
81.54% |
FFPE DNA-Mammary gland |
Rapid CE V2 |
2690282 |
99.45% |
48.66% |
98.02% |
94.15% |
99.99% |
81.90% |
Using Human blood gDNA and FFPE DNA as templates, RK20254 was used for library construction and Panel1 hybridization capture, and the capture efficiency of RK20254 could reach more than 70%.
Documents
Certificate of Compliance
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Lot number
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