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Advanced Techniques for ZF Recovery through RNA Interference and siRNA Design

Discover the pathway involving DICER and RISC, explore software for siRNA design, databases, and methods for siRNA delivery and testing. Design and test siRNA to suppress ZF efficiently.

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Advanced Techniques for ZF Recovery through RNA Interference and siRNA Design

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  1. Recovery of ZF by PCR FLAG+ FLAG-

  2. RNA interference May 9

  3. siRNA • discovery • pathway DICER and RISC • software for siRNA design • databases • delivery of siRNA • testing siRNA • designing and testing siRNA against ZF

  4. suppression of ZF by siZF 10ng 100ng 500ng pcZF psiZF#14 µg 0 0.5 1 0 0.5 1 0 1 0.5 1 0.5 1 psiRad9 µg 1 2 3 4 5 6 7 8 9 10 11 14

  5. U6 prom GFP mU6pro BbsI XbaI PolIII term BspI TTTG…………TTTT …………AAAAGATC XbaI si hairpin

  6. siRNA for ZF ATGGAGGAGGAGGCGATCGCCAGCCTCCCGGGGGAAGAGACGGAGGATATGGACTTTCTGTCTGGGCTGGAACTGGCGGATCTCCTGGACCCCAGGCAACCGGACTGGCACCTGGACCCCGGGCTTAGCTCGCCGGGGCCTCTCTCCTCGTCTGGCGGAGGCTCGGATAGCGGCGGCCTGTGGAGAGGGGACGATGACGATGAGGCCGCGGCTGCTGAAATGCAGCGCTTCTCTGACCTGCTGCAAAGGCTGTTAAACGGTATCGGAGGCTGCAGCAGCAGCAGTGACAGTGGCAGCGCCGAAAAGAGGCGGAGAAAGTCCCCAGGAGGAGGCGGCGGTGGCGGCAGCGGTAACGACAACAACCAGGCGGCGACAAAGAGTCCCCGGAAGGCGGCGGCGGCCGCTGCCCGCCTTAATCGACTGAAGAAGAAGGAGTACGTGATGGGGCTGGAGAGTCGAGTCCGGGGTCTGGCAGCCGAGAACCAGGAGCTGCGGGCCGAGAATCGGGAGCTGGGCAAACGCGTACAGGCACTGCAGGAGGAGAGTCGCTACCTACGGGCAGTCTTAGCCAACGAGACTGGACTGGCTCGCTTGCTGAGCCGGCTGAGCGGCGTGGGACTGCGGCTGACCACCTCGCTCTTCAGAGACTCGCCCGCCGGTGACCACGACTACGCTCTGCCAGTGGGAAAGCAGAAGCAGGACCTGCTGGAAGAGGACGACTCGGCGGGAGGAGTCTGTCTCCATGTGGACAAGGATAAGGTGTCGGTGGAGTTCTGCTCGGCGTGCGCCCGGAAGGCGTCGTCTTCTCTTAAAATGTAG #14 #15

  7. oligos

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