I like to leave them till there is lots of roots and they almost fill the flask. Depending on the size of flasks. I use the tall baby food jars,so when the leaves are near the top I take them out. If the leaves start turning brown I watch than take them out if they seam to have used up the nutrients in the agar. The longer they are in the flask the stronger they will be when taken out. this is only my opinon.