User:Karmella Haynes/Notebook/BioBrick cloning/2011/06/03

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06/03/11 ✓ Assemblies: CFP-Reporter and RFP-Repressor genes into MV8 (11) Assemblies KAH186/MV8: KAH186/(E/S)/2025 + MV8/(E/X)/4594 ✓ KAH184/MV8: KAH184/(E/S)/2191 + " KAH187/MV8: KAH187/(E/S)/2269 + " KAH188/MV8: KAH188/(E/S)/2269 + " KAH189/MV8: KAH189/(E/S)/2124 + " KAH190/MV8: KAH190/(E/S)/2368 + " KAH191/MV8: KAH191/(E/S)/2223 + " KAH183/MV8: KAH183/(E/S)/2383 + " KAH192/MV8: KAH192/(E/S)/2461 + " KAH196/MV8: KAH196/(E/S)/3212 + " KAH198/MV8: KAH198/(E/S)/3456 + " > Digests (Fermentas FD) --> Digest KAH196, 198 w/ E/S/ScaI Reagent Volume   30 μL/lane, 1% agarose DNA (plasmid) up to 25 μL 10x buffer 3.0 enzyme 1 1.0 enzyme 2 1.0 dH2O ---   30 μL --> 37°C/ ~30 min. > Measure conc.'s Sample OD260 260/280 ng/μL 1. KAH186 (E/S) 0.286 2.14 14.3 2. KAH187 (E/S) 0.176 2.58 8.8 3. KAH188 (E/S) 0.376 2.34 18.8 4. KAH189 (E/S) 0.209 2.36 10.4 5. KAH190 (E/S) 0.520 1.93 26.0 6. KAH191 (E/S) 0.326 2.16 16.3 7. KAH192 (E/S) 0.265 2.81 13.2 8. KAH196 (E/S) 0.669 2.01 33.5 9. KAH198 (E/S) 0.687 1.98 34.4 > Ligations Ligation Plate results (lig : neg crtl) 6/03/11 1. KAH186(E/S)/2025, 18 ng + MV8(E/X)/4594, 25 ng KAH186/MV8 >10:1 (Pick 2) 2. KAH184(E/S)/2191, 19 ng + " KAH184/MV8 >10:1 (Pick 2) 3. KAH187(E/S)/2269, 20 ng + " KAH184/MV8 >10:1 (Pick 2) 4. KAH188(E/S)/2124, 18 ng + " KAH184/MV8 >10:1 (Pick 2) 5. KAH189(E/S)/2296, 20 ng + " KAH184/MV8 >10:1 (Pick 2) 6. KAH190(E/S)/2368, 21 ng + " KAH184/MV8 >10:1 (Pick 2) 7. KAH191(E/S)/2223, 19 ng + " KAH184/MV8 >10:1 (Pick 2) 8. KAH183(E/S)/2383, 21 ng + " KAH184/MV8 >10:1 (Pick 2) 9. KAH192(E/S)/2461, 21 ng + " KAH184/MV8 >10:1 (Pick 2) 10. KAH196(E/S)/3212, 28 ng + " KAH184/MV8 >10:1 (Pick 2) 11. KAH198(E/S)/3456, 30 ng + " KAH184/MV8 >10:1 (Pick 2) 12. MV8(E/X)/ 25 ng     1-11 12 Insert DNA 3.5 --- Vector DNA 0.5 0.5 2x lgn buf (Roche) 5.0 5.0 T4 ligase (NEB) 1.0 1.0 dH2O --- 3.5   10 μL 10 μL