I am looking to use the Illumina DNA prep kit 96 well edition for metagenomics on stool extractions. I've heard that you can get many more samples then just 96 out of a single kit, how many can you actually get out based on the amount of buffers, beads, etc? Looking to see if anyone has empirical experience with this. Obviously the indexes will have to be on a second plate if there are more than 96. Thank you!
https://www.illumina.com/products/by-type/sequencing-kits/library-prep-kits/nextera-dna-flex.html
It would have been good if you also mention/refer the source you heard/got that information. Anyway, if the company develops any product, sure they include some extra volume of buffers etc. but that does not mean you can make more samples out of them. Additionally, if you use half the recommended volumes of reagent, you can simply double your output. No one will recommend that, but you can try and see/evaluate the results.
I think you are mixing two questions at a time, I will try to address them both.
1. Can we prepare the library for more than 96 samples.
Answer: Yes we can, it depends on the no of index primers in the kit and how many non-repetitive combinations you can have. Previously, we used to prepare 196 samples at a time using i.e 8 Forward and 12 reverse primers. So, Check your kit.
2. how many samples can be prepared using a kit, in terms of reagent volume.
Answer: Most of the time volume of Kapa Hi-fi or amount of polymerase enzyme is limiting, In our case, we easily used to prepare 196*2 samples using one bottle of the enzyme.
Caution: Don't change the volume of your regents, surely Its gonna affect your microbial ecology in post-sequencing output. Coz, it might be limiting for all the available DNA segments.
Thank you for the answers! It sounds like I will just use the prescribed amount then.
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