Figure 1: Dot Blot - Anti-N6-Methyladenosine Antibody [RM362] (A121405)
Dot blot of Lambda DNA without or with m6A, using Anti-N6-Methyladenosine Antibody [RM362]. The membrane was pre-spotted with 5 and 0.5 ng/dot of lambda DNA. m6A-free DNA was isolated from bacteriophage lambda grown in an E. coli host - deficient in adenine methylase (dam-).
MeRIP was performed using M6A and control RNA from NEB (E1610). Anti-N6-Methyladenosine Antibody [RM362] and Anti-PEG Antibody [RM105] were mixed for a short time, with Protein A beads before the addition of m6A labelled and non-labelled RNA (in equal amounts). Captured RNA was then washed, eluted, and run through RT-PCR. Resulting cDNA was then analyzed using real-time PCR with specific probes to m6A labelled and un-labelled sequences.