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Structural variations are present throughout the genome and are associated with various diseases. Mate-paired libraries, in conjunction with high-throughput SOLiD® next generation sequencing, provide a valuable method for detecting genomic rearrangements such as insertions, deletions, duplications, and translocations.

The SOLiD® Mate-Paired Library Construction Kits provide a fully optimized set of reagents for constructing mate-paired libraries for sequencing using the SOLiD® System.

Mate-paired Libraries

Mate-paired libraries are produced by randomly shearing DNA, size-selecting fragments, and bringing the fragment ends into close proximity by recircularization upon a synthetic oligonucleotide adaptor. Further manipulations produce a library of fragments that allows the sequence of both ends of each fragment to be determined sequentially. Aligned to a reference genome, the orientation and distance between the mate-pair sequences provide critical information for genomic assembly and identification of genomic variants.

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Figure 1: Schematic of SOLiD® Long Mate-Paired Library Construction Kit. Schematic of SOLiD® Long Mate-Paired Library Construction Kit (click to enlarge). Starting with sheared, size-selected gDNA, a final mate-paired library can be prepared as shown below. After ligating CAP Adaptors to the sheared DNA, the appropriate sized DNA is selected and circularized for P1 and P2 Adaptor ligation to beads and subsequent amplification.

Two mate-paired library construction methods are available. The SOLiD® Long Mate-Paired Library Construction Kit uses a protocol that captures ~50 bp on either end of the circularized fragment and uses a time-course S1 Nuclease digestion of the DNA (Figure 1). The SOLiD® 2 x 25 bp Mate-Paired Library Construction Kit uses a protocol that captures ~25 bp of either end of the circularized fragment and uses a restriction digestion with EcoP15 to generate the fragments (Figure 2). In general, capture of longer ends is preferred to provide as much information as possible and facilitate bioinformatic mapping. However, for several reasons including biases of each method, consistency of cuts, prior research, and organisms of interest, it may be desirable to generate libraries using both protocols.

The SOLiD® Mate-Paired Library Construction Kits:

  • Enable a convenient and simplified workflow — includes all reagents, columns, and gels for the construction of ten libraries (see note)
  • Optimized and recommended protocols for use with the SOLiD® System
  • Components quality-tested together to help ensure optimal results

Table 1

Product Part Number SOLiD® 2x25 Mate-Paired Enzyme Kit SOLiD® Long Mate Paired Enzyme Kits 1 & 2 SOLiD® Mate-Paired Bead & Buffer Kit SOLiD® Library Column Purification Kit SOLiD® Library Quick Gel Extraction Kit
SOLiD® 2x25 bp Mate-Paired Library Construction Kit4443472YesNoYesYesYes
SOLiD® Long Mate Paired Library Construction Kit4443474NoYesYesYesYes
SOLiD® Long Mate Paired Library Construction Reagents and Gel Extraction Modules4443711NoYesYesNoYes
Note: Does not include components of the SOLiD® Mate-Paired Library Oligos Kit (Cat #4400468), EcoP15I (used in the 2 x 25 bp protocol), or S-adenosylmethionine (used in the 2 x 25 bp protocol), which must be purchased separately. For research use only. Not intended for animal or human therapeutic or diagnostic use, unless otherwise indicated.

SOLiD® Software For Your Application

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