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Advanced Search Abstract RNA interference RNAi is a functional genomics tool to correlate genotype and phenotype by delivering targeted, gene-specific, and complementary dsRNA into a host via injection, feeding, or other means in order to reduce gene expression. Traditionally, successful transcript knockdown has been quantified by qPCR on a gene-by-gene basis, where only expression of the target gene and normalization genes are evaluated.

In this study, RNA-Seq was used to quantify transcript expression in larvae injected with dsRNA for aspartate 1-decarboxylase ADCwhich gives a reliable phenotype of an adult with a black cuticle instead of the wild-type red-brown. To evaluate the hypothesis that increased dopamine levels can affect mobility, T.

The data demonstrate that RNA-Seq can reveal gene connectivity and provide more complete data validation and analysis compared to qPCR.

RNA interference RNAi is a popular tool in functional genomics and, in pest insects like Tribolium castaneum, a potential strategy for molecular pest control Baum et al. RNAi is an evolutionarily conserved mechanism that relies on natural cellular pathways to target and degrade double stranded RNA dsRNA of viruses, as described in nematodes Fire et al. This natural process can be mimicked by the introduction of dsRNA into an organism e.

RNAi has been used successfully to increase insect mortality or developmental abnormalities via microinjection or feeding in vitro or in planta Baum et al. Other studies have demonstrated that injection can be successful in any T. This type of analysis is dependent on gene-specific primers, including onerous optimization that is often circumvented, and generally only used to measure the target gene and a normalizer gene as a standard.

For example, qPCR validation of knockdown of a major gut cysteine peptidase gene in T. However, using RNA-Seq, we demonstrated that knockdown was in fact successful, but closely related cysteine peptidase as well as serine peptidase genes, were compensating for the loss of target gene expression Perkin et al.

Thus, sequence similarity and redundancy in gene function, as well as compensation responses embedded in the transcriptome response, can mask the effects of RNAi, making it difficult to verify with qPCR alone.

The process is complex and requires the conjugation and cross-linking of cuticle proteins, leading to an insoluble, hard, and darkened red-brown exoskeleton Roseland et al. Knockdown of ADC leads to a black cuticle phenotype and the accumulation of dopamine Arakane et al.

One affected gene, dopamine receptor 2, led us to test whether movement was impaired in T.

Salaam Breaks Two OU Records, American Leader in the 60

Materials and Methods Insects The T. All primers were specific, and selected primer sets were unique to the ADC gene. The first round of PCR amplified the entire gene using the primary primer sets: A second set of primers was designed to amplify within the first amplification region, and the products were used for dsRNA amplification. Negative controls included noninjected larvae Control and mock-injected with water and dye Mock. Martin Invitational, Salaam has now PRed in the meter dash three straight times.

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The Big 12 champion in the meter dash, Salaam now owns Oklahoma indoor records in the 60 and meter dash. Saturday's performance also broke a two-year old meet record of 6. Salaam was not done for the day, however, as the junior returned to the track just over 30 minutes later to win the meter dash, breaking his own school record in the process with a time of The time broke his own school record of Salaam's time Saturday is the third fastest in the NCAA this year Shump won her second shot put title in as many weeks, hitting an NCAA automatic qualifying mark for the second straight week.

Shump's winning throw of Shump led the event from wire-to-wire, improving on each of her first three throws before hitting her winning throw on her third attempt.

Shump was one of two Sooners in the top three of the event as Tia Brooks finished third with a throw of Brittany Borman was fourth in the event with a personal best of The season has seen Mock compete in the long jump four times and come away with a win the last three times. Mock won her third title in as many weeks Saturday when she leapt The distance is just a half-inch off her season best of Like Mock, Smith won her third event title in as many weeks as she and Kristen Rice teamed to sweep the top two spots in the high jump.

Smith cleared the bar at Candyce McGrone improved her season time in the meter dash as she finished second in the event with a time of Scottesha Miller was third in the event with a time of Shelleyeka Rolle took second in the women's yard run with a time of 1: Like Rolle, Trayvion Jones improved his yard time as the sophomore finished third with a time of 1: Bill Kogel recorded his collegiate best in the mile as he finished second in the event with a time of 4: