VAHTS AmpSeq Library Prep Kit is based on ultra-multiplex PCR, employing several core techniques including end primer digestion, and adaptor ligation to construct amplicon libraries. The whole workflow is completed in one tube without need for cleanups. The minimum procedure time is 4 h from sample amplification to constructed libraries.
Size: 24 reactions.
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This kit enables construction of highly reliable and uniform libraries.
In combination with target amplification primer Panel, it enables highly efficient construction of libraries for target region. Specifically, in combination with VAHTS AmpSeq Cancer HotSpot Panel (Vazyme # NA102), it can be applied to prepare specific libraries for hotspot regions of the cancer-related genes.
For VAHTS AmpSeq Library Prep Kit, the initial template is 10 ng~100 ng, compatible with genomic DNA, FFPE samples, cell-free (cf) DNA and so on. It is compatible with Illumina and Ion Torrent high-throughput sequencing platform, in combination with VAHTS AmpSeq Adapters 1-12 for Illumina (Vazyme #NA110) or VAHTS AmpSeq Adapters 1-12 for Ion Torrent (Vazyme #NA120),
• One-tube: The process is simple, it takes only 4 hours from DNA to library generation, and the hands-on time is only about 30 minutes without the need for purification steps. Amenable to automation:
Figure Work Flow of VAHTS AmpSeq Library Preparation
• Compatibility with multiple types of sample: Compatible with genomic DNA, cfDNA, FFPE-derived DNA
• Very high on-target ratio exceeds 95%
• Excellent data uniformity: uniformity ≥95%
• Very low initial quantities of template: starting DNA template is as low as 10 ng, in some cases, 1 ng genome
• Contamination-resistant: End-primer digestion technology prevents libraries from being used as next-generation amplification templates
• Cost savings: Primer sequences are hardly sequenced, more sequencing data can be useful for your research
• Dual-platform compatibility: Compatible with the two mainstream platforms: Illumina and Ion Torrent
• Excellent amplicon GC compatibility: homogeneous and efficient amplification of different GC content amplicon
Results are greater than 95% On-target.
Figure 2: On-target proportions for sequencing different types of samples
Figure 2 shows the statistical results of the final data compiled from sequencing with the VAHTSTM AmpSeq Cancer HotSpot Panel (Vazyme #NA102). As can be seen from the figure, the on-target ratio is higher than 95% regardless of whether the starting template is derived from cell-extracting high-quality genomes or from complex templates such as FFPE samples and cfDNA. In addition, the repeated experiments confirmed excellent reproducibility.
Uniformity of data greater than 95%
Figure 3. On-target ratio of amplicons at different sequencing depths
Figure 3 shows the statistical results of the final results from sequencing with the VAHTSTM AmpSeq Cancer HotSpot Panel (Vazyme #NA102). As can be seen from the figure, the on-target ratio of amplicons at greater than 0.2 x mean depth exceeded 95%, and the on-target proportion of amplicons at greater than 0.5 x mean depth was greater than 90%. Excellent
Uniform and efficient amplification of amplicon with different GC content were demonstrated.
Figure 4. On-target ratio of multiple replicates from different samples
Figure 4 shows the statistical results of the down machine data of the VAHTSTM AmpSeq Cancer HotSpot Panel (Vazyme #NA102). As can be seen from the figure, the vast majority of amplicon sequencing with a GC content of 20% to 80% can reach or exceed the 0.2X average sequencing depth
For research use only. Not for use in clinical diagnostic or therapy.