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In vitro transcription with yeast nuclear extract
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摘要:In vitro transcription with yeast nuclear extract Steve HahnLast Modified Fri, Apr 25, 2003Wear gloves throughout, use RNAse free solutions (either autoclaved or sterile filtered) and clean bench and [阅读全文:]

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In vitro transcription
词条创建者:admin创建时间:12-09 22:51
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摘要:The following protocol is for MEGAscript II Kit (Ambion). 1. PCR amplify the DNA template. The 5'-end of the template should contain the minimum promoter sequenences of T7 or Sp6 or T3.A 5-primer with[阅读全文:]

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in?vitro?Transcription
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摘要:in vitro TranscriptionReaction1 µl 10X Transcription Buffer (Ambion) 1 µl 10X NTPs (4 mM ATP, CTP, 1 mM GTP, UTP) 2 µl 10 mM GpppG cap (Pharmacia) 2 µl a[32P]-UTP (NEN) 800 Ci/[阅读全文:]

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Transcription?of?cRNA?probe
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摘要:Transcription of cRNA probeReagents/SolutionsDNA (PCR or plasmid) with bacteriophage RNA polymerase promoter in suitable orientation (see note 1 ) ~1µg/µl in TE buffer 10mM MgCl2 5x Transc[阅读全文:]

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siRNA用户手册
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摘要:The siRNA user guide(revised May 6, 2004)Selection of siRNA duplexes from the target mRNA sequenceUsing Drosophila melanogaster lysates (Tuschl et al. 1999), we have systematically analyzed the silenc[阅读全文:]

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T7 siRNA protocol
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摘要:Here is our T7 siRNA protocol. The main idea is to design primers that begin with GG so thattranscription by T7 polymerase is efficient.I) FIRST design oligos:Using sense coding sequence of any gene..[阅读全文:]

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Protocol?for?First-strand?cDNA?Synthesis
词条创建者:admin创建时间:12-09 22:51
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摘要:Prepare in a sterile tube: template RNA:total RNA0.1-5µgor poly(A)+ mRNA10ng-0.5µg,or specific RNA0.01pg-0.5µg primer:oligo(dT)180.5µgor random hexamer 0.2µg,or sequence-[阅读全文:]

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siRNA protocols
词条创建者:admin创建时间:12-09 22:51
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摘要:Our current strategy with siRNA is to synthesis relatively small amounts enzymatically and use these to test for efficiency by western blotting. This allows us to test several candidate siRNAs quite c[阅读全文:]

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siRNAs的制备方法
词条创建者:admin创建时间:12-09 22:51
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摘要:越来越多的研究人员开始采用小分子干扰RNA(small interfering RNAs,siRNAs)来抑制特定的哺乳动物基因表达。siRNA是一种短片断双链RNA分子,能够以同源互补序列的mRNA为靶目标降解特定的mRNA,这个过程就是RNA干扰途径(R[阅读全文:]

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siRNA Design
词条创建者:admin创建时间:12-09 22:51
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摘要:RNAi target selection rules : Targeted regions on the cDNA sequence of a targeted gene should be located 50-100 nt downstream of the start codon (ATG). Search for sequence motif AA(N19 )TT or NA(N21 )[阅读全文:]

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RNA技术