User:Karmella Haynes/Notebook/BioBrick cloning/2011/01/25

{| width="800"
 * style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]] Project name
 * style="background-color: #F2F2F2" align="center"|  |Main project page
 * style="background-color: #F2F2F2" align="center"|  |Main project page


 * colspan="2"|
 * colspan="2"|

01/25/11

 * &#x2713; Minipreps: KAH160/pSB31 (10)
 * &#x2713; Cultures & minipreps: KAH165/pSB31 (10); 2 mL
 * &#x2713; Assemblies: KAH179, 180 (CFP-Targets) into KAH176, 177, 178/MV8 (RFP-Repressor expression vectors); continue later
 * &#x2713; Cultures: SP1AB 3x mut (24); 2 mL

Minipreps > Check with KpnI/NcoI digests --> pSB31 KpnI is located 142 bp upstream of EcoRI cloning site (according to pdf map) --> BioBrick constructs have 2 internal NcoI sites

--> Conclusions: KpnI digestion unclear; successfully identified KAH160/pSB31 clone; continue w/ picking more KAH165/pSB31 clones before sending plasmids in for sequencing. --> ID'ed two promising KAH165/pSB31 clones; send five putatives off for sequencing (1/26/11 Snow emergency: Genewiz not picking up orders until further notice)

Assemblies
 * 1) KAH179/176/MV8: (1) KAH179/(E/S)/2941 + (3) KAH176/MV8/(E/X)/7785
 * 2) KAH180/176/MV8: (2) KAH180/(E/S)/2743 + "
 * 3) KAH179/177/MV8: KAH179/(E/S)/2941 + (4) KAH177/MV8/(E/X)/7978
 * 4) KAH180/177/MV8: KAH180/(E/S)/2743 + "
 * 5) KAH179/178/MV8: KAH179/(E/S)/2941 + (5) KAH178/MV8/(E/X)/7989
 * 6) KAH180/178/MV8: KAH180/(E/S)/2743 + "

> Digests (Fermentas FD) --> Note: Re-digested KAH179 w/ E/S/ApaLI

> Measure conc.'s

--> Continue with ligation later


 * }