I am dephosphorylation RNA by Antarctic phosphatase and then decapping it by RppH (NEB). I am curious to know can i only use Rpph for for the process in one single reaction.As i have read RppH can also be used for dephosphorylation and decapping.
Hi there,
The NEB RppH enzyme is devoided of any detectable phosphatase activity as mentioned on the leaflet:
https://www.neb.com/~/media/Catalog/All-Products/1DBEEB2FECF54730AB4B0D17537537D1/Datacards%20or%20Manuals/M0356Datasheet-Lot0011305.pdf
Therefore you can't avoid using RppH and phosphatase (I don't know if you may use both of them in the same reaction... If not RppH comes first and then phosphatase) for total dephosphorylation of mRNA 5' end.
yes my manual also says the same but some other articles were reporting. Thanks for clearing me. I am doing 5' RACE so i need 5' monophosphate end to ligate my RNA with an adapter.
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