xGen¢ç Lockdown¢ç Probes Individually quality controlled capture probes for targeted sequencing xGen Lockdown Probes are individually synthesized and quality controlled 5¡Ç-biotinylated probes for hybridization capture target enrichment in next generation sequencing.
xGen Lockdown Probes
These probes can be used for creating custom capture panels that can be optimized, expanded, and combined with other panels. xGen Lockdown Probes can also be used to enhance the performance of existing capture panels, rescuing poorly represented regions such as areas of high GC content. With xGen Lockdown Probes, you can:
Preconfigured probe pools designed to target human gene coding sequences for cost-effective custom panels. xGen Predesigned Gene Capture Pools contain individually synthesized Lockdown¢ç Probes pre-formulated into individual gene pools for easy selection and setup of custom target panels.
xGen Predesigned Gene Capture Pools
xGen Predesigned Gene Capture Pools are preconfigured probe designs available for all human gene coding sequences. These probe pools are the best choice for creating cost-effective, custom gene panels that are delivered quickly. Probe designs are generated using the same design engine used to create the xGen Exome Research Panel, and use 1X tiling with repeat-masking. These probe sets have been demonstrated to provide high target region coverage and uniformity and minimal GC bias (see the Performance tab). xGen Predesigned Gene Capture Pools are designed using the GRCh37/hg19 reference genome and are available for any human RefSeq coding gene. Enter your target gene symbols into the xGen Lockdown Design Tool and receive your panel in 7–10 days. The pools are supplied either premixed in a single tube (16 or 96 reactions) or as individual gene pools delivered in 96-well plates (96 reactions only). With a minimum of 10 genes per pool, you can explore different panel compositions to identify the appropriate panel composition for specific projects. To include specific non-coding sequences in your target capture panel, simply add custom xGen Lockdown Probes.
High Target CoveragexGen Lockdown Probes provide approximately 5000-fold enrichment, high coverage over targeted regions (Figure 1), and minimal GC bias.
Figure 1. xGen Lockdown Probes Yield High Coverage. Ligation-based sequencing libraries were amplified by PCR using HiFi DNA polymerase (Kapa Biosystems). A set of 1000 xGen Lockdown Probes spanning the targeted region was used for in-solution hybrid selection with 2 µg of library input. Hybridization was performed for 24 hr and enriched targets were sequenced on a HiSeq 2000 System (Illumina) using 49 x 49 paired-end reads. The Lockdown Probes provided greater coverage than array/RNA baits. Improve Coverage of Array-Synthesized BaitsxGen Lockdown Probes can be added to array-derived RNA baits to enhance the coverage of GC-rich targets, such as first exons (Figure 2). Figure 2. Spiking In Individually Synthesized Oligonucleotide Baits Improves Sequencing Coverage of Array-Derived RNA Baits. Individually synthesized 5¡Ç-biotinylated Ultramer¢ç Oligonucleotides (xGen¢ç Lockdown¢ç Probes) were spiked into commercial, array-synthesized RNA baits to improve coverage of areas with high GC content. Either (A) 1000 Ultramer oligo baits targeting a ~133 kb region, or (B) 3 Ultramer oligo baits targeting a single exon, were mixed with 1.1 MB RNA baits and hybridized for 24 hr, followed by target capture using streptavidin. Enriched targets were sequenced on a HiSeq 2000 system using 49 x 49 paired-end reads. Lockdown probes improved general coverage of GC-rich targets (A) and single GC-rich exons (B). BrochuresAdditional reference protocols
Number of reactions and resuspension volumesInstructions:
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