Antigen selection of ARM complexes

Selection is carried out by adding the antigen-coated Dynabeads directly to the TNT translation mixture (Protocol 4). Non-binding complexes are removed by washing the beads. A magnetic particle concentrator (Dynal MPC) is used to wash and recover the beads canying bound ARM complexes. 

Preparation of antigen- or streptavidin-coupled magnetic beads 

Coupling of antigen to tosyl activated Dynabeads M-280 (see also manufacturer s Instructions) 

A convenient amount to prepare is 0.5-1 mg (50-100 pil) beads. Wash beads with 0.5-1 ml buffer A, 

Mix with antigen in proportions of 20 jLgantigen l mg beads and vortex for 1 min. Incubate for 16-24 h at 37 °C with slow rotation. 

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Construction of fiagments from hybridomas and lymphocyte mRNA 95 Generation of ARM complexes by coupled transcription/translation in vitro 97 Preparation of antigen- or streptavidin-conpled magnetic beads 98 Antigen selection of ARM complexes 99... 

Recovery and amplification of DNA from antigen-selected ARM complexes... 

Recovery and amplification of genetic infonnation from antigen-selected ARM complexes Screening of clones by ARM displ using an internal standard in RT-PCR 104... 

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