This investigation was directed to characterizing 30 Assam Lemon accessions based on morphological and biochemical characters. The dissimilarity matrix ranged from 4.72 to 11.60 indicating diverse nature of collected accessions. The lowest coefficient of dissimilarity (4.72) was observed between KP-1 and TZ-3 signifying that they were closely related. The highest coefficient of dissimilarity (11.60) was observed between TZ-1 and KN-2 signifying that they were the most distant of all the collected accessions. Dendogram based on the Euclidean distance using UPGMA cluster analysis of morphological and biochemical characters, differentiated 30 accessions in to three major clusters, cluster-I, cluster-II and cluster-III. Thus, cluster analysis revealed sufficient variation between the collected accessions by grouping them into different clusters. The principal component analysis (PCA) of the traits revealed that the first 12 principal components were important out of total 30 components and they accounted for 82.71% of the total variability. Based on PCA and correlation matrix of 34 morphological and biochemical traits, 23 traits were identified for easy evaluation and preliminary characterization of Assam Lemon germplasm. The clustering pattern based on 34 traits and that based on 23 traits were found to be identical indicating that the selected traits were likely to cover the existing diversity of the variety.