The Knicks know Mozgov is long (7 feet 1 inch, 250 pounds) and athletic, a decent shooter and a competent shot-blocker.
After ligation, the DNA was transformed into chemically competent One Shot Top10 Escherichia coli (Invitrogen, Carlsbad, CA, USA).
Amplicons were ligated en masse into the Topo TA cloning vector (Invitrogen) and transformed into competent one shot Topo top 10 bacterial cells (Invitrogen).
Two microliters of undiluted ligation mixture were added to 25 mls of chemically competent One Shot® Top10 E.coli cells (Invitrogen, Carlsbad, CA, USA) and mixed gently.
Plasmids including the DNA fragments of interest were transformed into the chemically competent One Shot TOP10 cells provided by the kit, according to the manufacturer's instructions.
The PCR products were cloned into plasmid vector pCR4-TOPO and transformed into competent One Shot TOP10 cells with a TOPO TA cloning kit (Invitrogen) following the supplier's protocol.
The liagtion mixtures were used either directly for PCR (referred from now as Lig-PCR) or for transformation of chemically competent One Shot® Top10 E.coli cells (Invitrogen, Carlsbad, CA, USA).
Cloning reaction products were used to transform chemically competent One Shot® TOP 10 E. coli cells (Invitrogen), and miniprep DNA from transformants were digested with restriction enzymes (MBI Fermentas) and analyzed by agarose gel electrophoresis to verify the presence of the gene of interest.
For plasmid production, plasmid was transformed into the chemically-competent One Shot Stbl3 E. coli (Invitrogen, Carlsbad, CA) following the manufacturer's protocol, isolated, and purified for genome-specific sequencing to confirm the presence of the desired insert.
PCR products were cloned using the TOPO TA Cloning Kit for subcloning with electro-competent One Shot TOP10 Escherichia coli cells (Life Technologies).
Lipofectamine 2000, mouse monoclonal V5 antibody, fluorescent labeled antibodies, the pcDNA3.1D cloning kit, E. coli One Shot competent cells, the pENTR Directional TOPO cloning kits and the Gateway mammalian expression system were from Invitrogen (Carlsbad, CA, USA).
