So I have been trying to amplify a deletion in the HbA1 and HbA2 genes (the genes that code for alpha domains in hemoglobin) and so far I've had no luck. The region is about 70% GC rich, and my amplification products (depending on the primers I've used) have ranged from 2100 bp to 1000 bp. I've tried adding DMSO and betaine. No luck. I used a Qiagen kit with something called "Q solution" (pretty much betaine) and used an exact protocol somebody else used with the same primers and had no luck. I've decreased annealing temp down to 48 and increased it to 60. I've increased initial denaturation to 95 C for 15 min. In total I've run about 20 reactions and not once got a band at the correct length, and mostly got no band at all.