What limitation did newer, culture-independent sequencing techniques overcome compared to 16S rRNA gene sequencing?

Capturing a more complete picture of microbial function and diversity.

Early sequencing methods, such as 16S rRNA gene sequencing, were valuable because they provided snapshots indicating which microbial species were present in a sample. However, these methods often proved insufficient for understanding the operational capacity of the community. Newer, culture-independent techniques were developed specifically to move beyond simple identification. They allow scientists to capture data related to the functional capacity of the microbiome—how the microbes are actually behaving, what metabolic pathways they are using, and the overall breadth of different types of organisms present—which is crucial for modern tools aiming to translate biological data into health insights.