Characterization of Cyanobacteria in lakes with different physicochemical properties provides insights into the diversity of this phylum and knowledge of their features that are relevant to biotechnology applications. Six Cyanobacterial isolates were recovered from freshwater Lake Nasser and saline Lake Qarun, Egypt. The isolates were identified based on both morphology and molecular markers, 16S rRNA, and RuBisCOcbbLgenes. The isolates SN1, SN2, SN3, SN4, Q1, and Q2 showed homologies withMerismopedia,Oscillatoria,Limnothrix,Persinema, andJacksonvillea, respectively. ThecbbLsequences for isolates SN1, Q1, and Q2 represented the first records for candidates relating to the generaMerismopediaandPersinema, andJacksonvillea, respectively. Biochemical contents, carbohydrates, proteins, lipids, pigments, and ash-free dry weight were measured for each isolate. Isolate SN2 had the highest content of allophycocyanin, 71 ± 4.8 mg/g DW, and phycoerythrin, 98 ± 6.7 mg/g DW, while the isolate SN4 had the highest composition of total protein, lipid, carotenoid, and chlorophyll a, recording 364.7 ± 6.4 mg/g DW, 67.6 ± 0.2 mg/g DW, 0.261 ± 0.01 mg/g DW, and 10 ± 0.6 mg/g DW, respectively. Isolate Q1 recorded the maximum amount of phycocyanin, 114 ± 20.7 mg/g DW among isolates. The isolate Q2 was observed to have the highest carbohydrate content, 274 ± 14.5 (mg/g DW), and ash-free dry weight, 891.8 ± 2.8 mg/g DW. Thus, the study indicated that the current isolates may represent promising resources for biotechnological applications.