Identification, Characterization, and Expression Analysis of Immunoglobulin Genes from Antarctic Fish by PCR Methods

We describe different applications of the polymerase chain reaction (PCR) method to amplify up to 2.5-kb DNA regions of the immunoglobulin genes from Antarctic teleost fishes and to evaluate their expression in different tissues. To enhance amplification of poorly expressed transcripts or difficult to amplify, for example, products generated by 5' or 3' RACE, we have set up optimal primer annealing conditions based on a three-step cycle strategy.The protocols reported here along with materials required and general tips are modifications of conventional PCR.