Detection Method for Multiple Gene Expression in Single Early Bovine Embryo (Antisense RNA-RT-PCR).
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- Kadokawa Hiroya
- Hokkaido National Agricultural Experiment Station
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- Hamano Seizo
- Animal Bio-Technology Center, Livestock Improvement Association of Japan
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- Itoh Ryuji
- Hokkaido-branch, National Institute of Animal Health
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- Takahashi Hitomi
- National Institute of Animal Industry
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- Yamada Yutaka
- Hokkaido National Agricultural Experiment Station
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- Kariya Takayoshi
- National Institute of Animal Industry
Bibliographic Information
- Other Title
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- ウシ初期胚内における複数の遺伝子発現に関する検討(アンチセンスRNA-RT-PCR法)
- ウシ ショキ ハイナイ ニ オケル フクスウ ノ イデンシ ハツゲン ニ カン
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Abstract
We examined whether we can detect multiple gene expressiom in single early bovine embryos by using antisense RNA (aRNA) and RT-PCR (aRNA-RT-PCR). Embryos, from the 2-cell stage to the blastocyst stage, were produced by the IVM-IVF method, cryopreserved and used as material for aRNA synthesis. Antisense RNA solution was yielded after reverse-transcription (RT) with a synthetic primer containing the T7 RNA polymerase binding site, double strand cDNA synthesis and transcription by the T7 RNA polymerase. For the reverse transcription of aRNA (aRNA-RT), we used a 5 μl of total 100 μl aRNA solution (1/20) and either sense primer or an antisense one encoding bovine beta-actin (bActin). After the aRNA-RT and PCR, the presence of the expected products was confirmed by electrophoresis. We confirmed the bActin products when the sense primer was used in aRNA-RT, but we did not confirm the products with the antisense one. These results indicate that we could detect multiple gene expression in a single early bovine embryo by means of aRNA-RT-PCR.<br>
Journal
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- Journal of Mammalian Ova Research
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Journal of Mammalian Ova Research 13 (2), 125-129, 1996
JAPANESE SOCIETY OF OVA RESEARCH
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Details 詳細情報について
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- CRID
- 1390001204340334080
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- NII Article ID
- 10013833522
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- NII Book ID
- AN10548943
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- ISSN
- 13475878
- 13417738
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- NDL BIB ID
- 4078627
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- Text Lang
- ja
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- Data Source
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- JaLC
- NDL
- Crossref
- CiNii Articles
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- Abstract License Flag
- Disallowed