---

For the convenience of users, particularly for the vast majority of experimental scientists, a publicly accessible web-server: H2Opred has been established. Moreover, to maximize users' convenience, a step-by-step guide is given below on how to use the web-server and get the desired results without the need to through the mathematical details of the algorithm.

• Step 1
  Go to the H2Opred homepage by copying the following link https://balalab-skku.org/H2Opred/ to the web browser or click here, and you will see its top page of H2Opred. Click on the Help button to see a brief introduction about the server (this page).

• Step 2
  We have developed the H2Opred 2'-O-methylation (2OM) nucleotide-specific prediction models and generic model for four different 2OM sites. Therefore, users can select appropriate 2OM sites for the prediction using the dropdown menu.

• Step 3
  Paste your query RNA sequences into the input box or directly upload your input file via the "Choose File" button. The input sequences must be in the FASTA format otherwise the program will throw an error. We have provided a set of sample FASTA sequences. Users can click (Ex: N2OM), (Ex: A2OM), (Ex: C2OM), (Ex: G2OM), or (Ex: U2OM) for easy testing of the H2Opred server.

• Step 4
  By clicking on the submit button, you will get the prediction results on your computer screen.
  • For the selected 2OM nucleotide-specific model, four columns are displayed. The first, the second, the third, and the fourth columns represent serial number, RNA id (i.e. user FASTA ID), 2OM or Non-2OM and probability of prediction.
  • Notably, the probability score ranges from 0 to 1, where a score close to 0 or 1 represents Non-2OM or 2OM, respectively.

• Step 5
  Click on the Reset/Clear button if you need to erase the query sequences present in the input box or file upload button.

• Step 6
  Click on the Download button to download datasets (training and independent) used in this paper.