Cell Receptor Discovery Protocol

1. Isolate RNA from a given cell

2. Use oligodT sepharose to isolate mRNA through hybridization with polyA tail of the mRNA

3. Make a labelled cDNA from the mRNA from the cell, 32P-labelled cell cDNA

4. Subtract 32P-T cDNA with unlabelled cell polyA and RNA. Complementary sequences will hybridize and separate single from double stranded material.

5. The subtracted 32P-labelled probed is subsequently used to probe a similarily made unlabelled cell cDNA library.

6. Verification of clones through Northern Blot. Label the isolated cDNA through the use of probe cells. RNA from the cells is run on a gel and then transferred to nitrocellulose and the nitrocellulose is probed with labelled cDNA for hybridization with complementary sequences.