Target-Enriched Multiplexed Parallel Analysis For Assessment Of Risk For Genetic Conditions

A method of determining the status of a genetic condition, the method comprising:

• (a) preparing a sequencing library from a sample of DNA;
• (b) hybridizing the sequencing library to a pool of TArget Capture Sequences (TACS), wherein the pool of TACS comprises a plurality of sequences that bind to genomic sequences of interest including variant allele loci of interest associated with different genetic conditions, wherein:
• (i) each member sequence within the pool of TACS is between 100-500 base pairs in length, each member sequence having a 5′ end and a 3′ end;
• (ii) each member sequence binds to the same genomic sequence of interest at least 50 base pairs away, on both the 5′ end and the 3′ end, from regions harboring Copy Number Variations (CNVs), Segmental duplications or repetitive DNA elements;
• and
• (iii) the GC content of the pool of TACS is between 19% and 80%, as determined by calculating the GC content of each member within the pool of TACS;
• (c) isolating members of the sequencing library that bind to the pool of TACS to obtain an enriched library;
• (d) amplifying and sequencing the enriched library;
• and performing statistical analysis on the enriched library sequences to thereby determine the carrier status at the loci of interest associated with different genetic conditions.

A kit comprising:

• a) a container comprising the pool of TACS, wherein the pool of TACS comprises a plurality of TACS families, wherein each TACS family comprises a plurality of member sequences, wherein each member sequence binds to the same genomic sequence of interest but has different start and/or stop positions with respect to a reference coordinate system for the genomic sequence of interest, and further wherein:
• (i) each member sequence within each TACS family is between 100-500 base pairs in length, each member sequence having a 5′ end and a 3′ end;
• (ii) each member sequence binds to the same genomic sequence of interest at least 50 base pairs away, on both the 5′ end and the 3′ end, from regions harboring Copy Number Variations (CNVs), Segmental duplications or repetitive DNA elements;
• and
• (iii) the GC content of the pool of TACS is between 19% and 80%, as determined by calculating the GC content of each member within each family of TACS and
• b) instructions for using the pool of TACS in a method of determining the status of a genetic condition.