Purpose: The aim of this study was to investigate and identify the rules of marking and punctuation in the selected dated manuscript in the library of Astan Quds Razavi and some other libraries. Methodology: This applied research was conducted through an analytical survey method. By reviewing the manuscripts in Astan Quds Razavi and after consulting with some experts working in this section, 6 dated manuscripts related to the considered subject and period as well as 6 manuscripts from other valid libraries were selected using purposeful sampling method. Results: By systematically examining and categorizing the signs used in selected manuscripts until the Timurid era, it was found that the sign (grear spot) and the sign "letter ﻫ" each with 41%, "three-point- triangle and three-point diamond” with 34%, were most used among the pictorial signs. Among the most widely used text signs with the letters in the manuscripts, "strain letters" with 50%, "spacing" and "vermilion writing" with 40% were at top. In 76% of selected manuscripts, the both types of signs were used. Conclusion: This study showed that the forms of "visual cues" and "signs with text letters Edition were used for marking in manuscripts. All these signs were used for separation and distinction and in most manuscripts both signs were used.