User guide

Experimental flowchart for single-subunit protein expression
Experimental
flowchart for
single-subunit
protein expression
The diagram below schematically depicts the steps necessary to express your
single-subunit protein of interest using the Freedom
™
DG44 Kit as well as several
common pathways from stable transfectants to clone scale-up. Note that the times
shown for various experimental steps are approximations, and the actual times
will depend on your protein of interest and the specific workflows you choose.
See Methods for Single-subunit Protein Expression, pages 42–61, for detailed
protocols.
Note: For detailed information on cloning your protein of interest into the
pOptiVEC
™
-TOPO
®
TA vector, refer to the pOptiVEC
™
-TOPO
®
TA Cloning Kit
manual (part no. 25-0977), available at www.lifetechnologies.com/manuals.
Important: If you are expressing a single-subunit protein, you must generate your
expression construct using the pOptiVEC
™
-TOPO
®
TA vector. The pOptiVEC
™
expression construct allows genomic amplification by MTX amplification.
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