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231223s2007 xx ||||| 00| ||eng c |
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|a pubmed25n0623.xml
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|a (DE-627)NLM186831889
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|a (NLM)19259483
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|a DE-627
|b ger
|c DE-627
|e rakwb
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| 041 |
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|a eng
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| 100 |
1 |
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|a Sukno, Serenella A
|e verfasserin
|4 aut
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| 245 |
1 |
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|a Quantitative Detection of Double-Stranded RNA-Mediated Gene Silencing of Parasitism Genes in Heterodera glycines
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| 264 |
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|c 2007
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| 336 |
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|a Text
|b txt
|2 rdacontent
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| 337 |
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|a ohne Hilfsmittel zu benutzen
|b n
|2 rdamedia
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|a Band
|b nc
|2 rdacarrier
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|a Date Completed 25.09.2012
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|a Date Revised 16.03.2024
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|a published: Print
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|a Citation Status PubMed-not-MEDLINE
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|a The introduction of a double-stranded RNA (dsRNA) into an organism to induce sequence-specific RNA interference (RNAi) of a target transcript has become a powerful technique to investigate gene function in nematodes and many organisms. Data provided here indicate that the inclusion of 1-2 mM spermidine and 50 mM octopamine and a 24 hr incubation period of nematodes in double-stranded RNA (dsRNA) soaking solutions resulted in a considerable increase in the percentage of nematodes that ingested dsRNA as compared to previous reports. This modified dsRNA soaking method was coupled with quantitative real-time RT-PCR (qRT-PCR) analyses to assess the potential silencing of the Heterodera glycines parasitism gene transcripts Hg-pel-1 and Hg-4E02 that are expressed within the esophageal gland cells of preparasitic H. glycines J2. The Hg-pel-1 transcript was most efficiently silenced with one dsRNA construct (ds267) at the highest dsRNA soaking concentration of 5.0 mg/ml, while the Hg-4E02 transcript was more efficiently silenced at the 2.5 mg/ml dsRNA concentration as compared to 5.0 mg/ml. A dsRNA construct (ds285) complementary to a different sequence within the Hg-pel-1 transcript than construct ds267 induced only minimal silencing of the Hg-pel-1 transcript at 2.5 mg/ml. The results suggest that both dsRNA concentration and sequence relative to the transcript targeted are critical for maximizing potential RNAi effects in parasitic nematodes
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|a Journal Article
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1 |
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|a McCuiston, Jamie
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Wong, Mui-Yun
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Wang, Xiaohong
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Thon, Michael R
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Hussey, Richard
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Baum, Thomas
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Davis, Eric
|e verfasserin
|4 aut
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| 773 |
0 |
8 |
|i Enthalten in
|t Journal of nematology
|d 1969
|g 39(2007), 2 vom: 28. Juni, Seite 145-52
|w (DE-627)NLM098196421
|x 0022-300X
|7 nnas
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| 773 |
1 |
8 |
|g volume:39
|g year:2007
|g number:2
|g day:28
|g month:06
|g pages:145-52
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|a AR
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| 952 |
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|d 39
|j 2007
|e 2
|b 28
|c 06
|h 145-52
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