What is Agilent SureSelect?
SureSelectXT Reagent Kits The SureSelectXT Reagent Kits provide DNA library prep solutions with hybrid capture-based target enrichment. They include all reagents for library preparation, hybridization, and capture. SureSelectXT Reagent Kits enable post-capture pooling of enriched, sequencing-ready libraries.
What is HaloPlex?
The HaloPlex protocol is optimized for digestion of 200 ng of genomic DNA (split among 8 different restriction digestion reactions) plus 25 ng excess DNA, for a total of 225 ng genomic DNA. Using lower amounts of DNA in the enrichment protocol can adversely affect your results.
What is Hybrid Capture?
Overview. Hybridization capture is a targeted next generation sequencing method that uses long, biotinylated oligonucleotide baits (probes) to hybridize to the regions of interest. It is particularly helpful when genotyping, detecting rare variants, and exome sequencing.
What can exome sequencing do for you?
ExomeSeq is a test that looks at most of the genes. This test may be useful for patients whose medical and family histories suggest a genetic cause for their signs and symptoms. Most patients who have whole exome sequencing (WES) have had other genetic testing that did not find a genetic cause of their condition.
What are the limitations of whole exome sequencing?
The major limitation of exome sequencing may be the inability to comprehensively represent genomic SVs. Many groups have designed algorithms that use a read depth or read pair-based approach for predicting structural variation; however, these approaches are not very efficient at identifying SVs with exome data.
What is target capture?
Targeted capture or target enrichment allows for simultaneous sequencing of sets of genomic regions of interest in several individuals in the same sequencing run, therefore reducing sequencing costs. This technique uses custom RNA probes which are complementary to the target genomic regions.
When should I order whole exome sequencing?
WES can be done in pregnancy, childhood, or adulthood. It is usually done for one or more of the following reasons: You are (or your child is) suspected of having a genetic condition. There are many different genes that can cause your (or your child’s) symptoms.
Which of the following is a disadvantage of exome sequencing?
Which of the following is a disadvantage of exome sequencing? Exome sequencing only identifies conditions associated with recessive alleles. Exome sequencing does not directly identify the gene, but identifies only the region of the genome containing the gene.
What can exome sequencing not detect?
Exome sequencing is limited in detecting the following types of mutations (this list might not be exhaustive): large rearrangements. copy number variation mutations (large deletions/duplications) mitochondrial genome mutations.
How do sureselect and haloplex work?
Target enrichment workflows of SureSelect and HaloPlex integrate the TapeStation and Bioanalyzer systems for quantitative and qualitative assessment of samples at various key steps (Figure 1). Materials and methods
What reagent kits did Agilent Technologies provide for DNA extraction?
(G9681A), HaloPlex (G9901C), and HaloPlexHS(G9931C) reagent kits were obtained from Agilent Technologies. Quantity determination of DNA libraries was measured using the region functionality.
What is the size shift between sureselectxt and sureselect XT2?
At this step, a size shift of approximately 60–80 bp with SureSelectXT(Figure 5) and 80–100 bp with SureSelectXT2(Figure 6) with respect to post sheared library is expected.
What are the four main QC points in the sureselectxt library preparation workflow?
SureSelectXTand SureSelectXT2 Multistep adaptor ligation library preparation methods such as SureSelectXTand SureSelectXT2have four main QC points in the overall library preparation and capture workflow: input DNA, post shearing, amplified precapture library, and amplified post capture library (Figure 1A)1.