BerkiGEM2007-AustinConstructionFile25

From 2007.igem.org

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Beta portion<br>
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Alpha: <br>
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pcr CA1154F/G00101 on 9145-I716063 (575 bp)<br>
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PCR CA1150/CA1099R on I716077 (1727 Bp)<br>
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Product is ADtemp1b<br>
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Digest (Bsa1/Bgl2) and ligate with beta portion.<br> <br>
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Sub ADtemp1b into pTRC plasmid with (Bgl2/XhoI)<br>
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Product is ADtemp2b<br>
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Beta: <br>
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pcr CA1099R/D58 on ADtemp2b (3465 bp)<br>
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Product is ADtemp3b<br>
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PCR CA1099R/D58 on Part Ptrc-I716082 <br>
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_______________________________________________________<br>
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Digest (Bsa1/BamH1) and ligate with alpha portion. <br>
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Alpha portion<br>
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<br><br>
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Pcr CA1150F/CA1099R on I716077 ()<br>
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Product is pTRC-I716075 (Best all around)
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Product is ADtemp5b<br>
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____________________________________________________________<br>
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Digest ADtemp3b (BsaI/BamHI)<br>
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Digest ADtemp5b (BsaI/BglII)<br>
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Ligate, product is: I716075 (Best all around)
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<br>
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Revision as of 21:50, 2 July 2007

Alpha:
PCR CA1150/CA1099R on I716077 (1727 Bp)
Digest (Bsa1/Bgl2) and ligate with beta portion.

Beta:

PCR CA1099R/D58 on Part Ptrc-I716082
Digest (Bsa1/BamH1) and ligate with alpha portion.


Product is pTRC-I716075 (Best all around)