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ive read where on some boards if you use the top slot for nvme it will steal lanes from the GPU but is it only Gen 5 drives that do that? i will be sticking with a mix of Gen 3 and 4 but i wasnt sure on that part. ive got a B650E board.
Consult the manual. ALL boards are different. Usually the top NVME x4 is fed directly from the CPU on modern platforms and doesn't interfere, but still consult the manual to verify this. Some NVME M.2 slots are fed by CPU, some are fed by chipset.
Sometimes when you're populating other NVME slots, it can cause other things to happen- like your primary x16 GPU slot drops down to x8, you lose some SATA ports, etc... All verified in the motherboard manual (including what generation slot is is e.g. gen 4 or gen 5)