User:Karmella Haynes/Notebook/BioBrick cloning/2009/12/29

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12/29/09

 * &#x2713; Minipreps: KAH136(2), 137(4), 138(4); retransformations KAH93(2), JDS42(2), JDS43(2)
 * &#x2713; Assemblies: KAH139, 140; KAH117-119; KAH94, 95/JDS42; KAH94, 95/JDS43
 * &#x2713; Cultures: KAH137, 138 (6 each)

Minipreps > Check with E/P digests

--> 37°C/ ~15 min. --> KAH137, 138 failed; pick 6 more colonies each

Assemblies
 * 1) KAH139: (1) KAH135/(E/S)/2353 + (3) NLSstop2(KAH011)/(E/X)/~3200
 * 2) KAH140: (2) KAH136/(E/S)/1941 + "
 * 3) KAH117: KAH99/(E/S)/2913 &#x2713; + (4) KAH93/(E/X)/8039
 * 4) KAH118: KAH100/(E/S)/7930 &#x2713; + "
 * 5) KAH119: KAH101/(E/S)/8002 &#x2713; + "
 * 6) KAH94/JDS42: KAH94/(E/P+Ad's)/5011 &#x2713; + (5) JDS42/(BglII)/7468
 * 7) KAH95/JDS42: KAH95/(E/P+Ad's)/5083 &#x2713; + "
 * 8) KAH94/JDS43: KAH94/(E/P+Ad's)/5011 &#x2713; + (6) JDS43/(BglII)/7478
 * 9) KAH95/JDS43: KAH95/(E/P+Ad's)/5083 &#x2713; + "

> Digests (Fermentas FD) --> BglII single digest for JDS42, 43 (2.0 μL BglII)

> Measure conc.'s

> Ligations --> Use dsoligo adapters (Ad BglII/EcoRI and Ad PstI/BglII from 12/28/09) for JDS constructs

--> For reactions 8-13, add everything except the vector DNA, R.T./ 10 min.; then add vector, R.T. 10 min. --> Add 20 μL Z-DH5α


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