Hey all,
I'm curious if there is any downside to using Herculase as opposed to the Illumina kit DNA polymerase for the final amplification of a library prep (after size selection). I ask because it's cheap, has worked well for me with other things, and in the Agilent SureSelect capture protocols it is used for the post-capture amplification, so I don't have reason to think it would cause any weird biases. I also tried using Herculase to see if I could get good amplification and I believe I can actually get enough product with fewer cycles than I could with the kit DNA polymerase (I haven't tested this thoroughly but I have reason to think this). I did this using the PCR protocol for post-capture amplification. Am I missing anything or can I simply use Herculase without having to worry about anything else?
I'm curious if there is any downside to using Herculase as opposed to the Illumina kit DNA polymerase for the final amplification of a library prep (after size selection). I ask because it's cheap, has worked well for me with other things, and in the Agilent SureSelect capture protocols it is used for the post-capture amplification, so I don't have reason to think it would cause any weird biases. I also tried using Herculase to see if I could get good amplification and I believe I can actually get enough product with fewer cycles than I could with the kit DNA polymerase (I haven't tested this thoroughly but I have reason to think this). I did this using the PCR protocol for post-capture amplification. Am I missing anything or can I simply use Herculase without having to worry about anything else?