What amplification method, if required, is recommended after the immunoprecipitation?
We recommend T7-based Linear Amplification of DNA (TLAD). TLAD begins with a terminal transferase (TdT) tailing of the template DNA, is followed by annealing with a primer adapter containing the T7 promoter sequence. A second strand is synthesised with DNA Polymerase I Klenow fragment, and then in vitro transcription of the template generates a RNA amplification product (Liu et al, 2003). This results in linear rather than exponential amplification, and this method has the potential to reduce potential bias introduced by PCR-based amplification.
