HomeFAQsI did not get enough yield of RNA for my downstream application after two rounds of IP. How do I increase the yield?
FAQ: I did not get enough yield of RNA for my downstream application after two rounds of IP. How do I increase the yield?
One way to increase recovery of RNA is to do one round of IP instead of two. One round typically generates enough enrichment of m6A containing RNA. Alternatively, the IP can be done with 2, 3, or 4 µl of antibody per round for 1 or 2 rounds. The use of extra antibody will result in more RNA to be recovered and the amount of enrichment will still be significant. However, use of >4 µl antibody might cause the enrichment to go down because a significant amount of unmodified RNA will also be recovered.