Wiki/Team:Warsaw/igem notebook.htm

Our protocols

• Purification of His_Z_alpha and His_Z_omega
• Purification of His_A_alpha
• Testing various hunter/prey combinations
• Plasmid DNA isolation
• DNA isolation from agarose gel
• DNA purification after enzymatic reaction
• Genomic DNA isolation
• Preparation of chemocompetent bacteria
• Preparation of electrocompetent bacteria
• Chemotransformation
• Electrotransformation
• Ligation
• Blunting
• Standard concentrations of antibiotics and other supplements
• Rifampicin test
• Removing 5' phosphate groups from DNA ends
• TAXI protocol (Tet+Ap 30+X-Gal+IPTG)
• Protein concentration measurement (BCA method)
• DNA digest
• PCR
• Polymerase Chain Ligation (PCL)

If not stated otherwise, all agarose gels bear Fermentas GeneRuler (100-10000 bp) ladder. PAGE gels bear PageRuler Fermentas Prestained protein ladder.

Our primers

Our vectors

Our strains

Rosetta(DE3)pLysS

F- ompT hsdSB(RB- mB-) gal dcm λ(DE3 [lacI lacUV5-T7 gene 1 ind1 sam7 nin5]) pLysSRARE (CamR)

TOP10 (Invitrogen)

F- mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 nupG recA1 araD139 Δ(ara-leu)7697 galE15 galK16 rpsL(StrR) endA1 λ-

NR9458 (IBB collection; in notebook called mutD5)

ara thi Δ pro lac mutD5 zaf-13 ::Tn10