BerkiGEM2007-AustinConstructionFile24

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-I716072 (575 bp)<br>
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PCR CA1150/CA1099R on 9145-I716077 (1727 Bp)<br>
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Product is ADtemp1<br>
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Digest (Bsa1/Bgl2) (2111/Small, Larger) and ligate with beta portion.<br> <br>
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Sub ADtemp1 into pTRC plasmid with (Bgl2/XhoI)<br>
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Product is ADtemp2<br>
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Beta: <br>
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pcr CA1099F/D58 on ADtemp2 (3465 bp)<br>
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Product is ADtemp3<br>
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PCR CA1099F/G00101 on Part 9145-I716081 <br>
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_______________________________________________________<br>
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Digest (Bsa1/BamH1) (1447/small, Larger) 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 9145-I716074 (best yield)
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Product is ADtemp5<br>
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____________________________________________________________<br>
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Digest ADtemp3 (BsaI/BamHI)<br>
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Digest ADtemp5 (BsaI/BglII)<br>
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Ligate, product is: I716074 (Best yield)
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<br>
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Latest revision as of 22:45, 10 July 2007

Alpha:
PCR CA1150/CA1099R on 9145-I716077 (1727 Bp)
Digest (Bsa1/Bgl2) (2111/Small, Larger) and ligate with beta portion.

Beta:

PCR CA1099F/G00101 on Part 9145-I716081
Digest (Bsa1/BamH1) (1447/small, Larger) and ligate with alpha portion.


Product is 9145-I716074 (best yield)