Q1: What samples are compatible with this kit?
Purified total DNA from any organism free from PCR inhibitors. This system has been used by Zymo Research’s Microbiomics Services team to process samples from human, water, soil, food, plants, feces, biofilms, and much more.
Q2: What is the minimum DNA input for this kit?
The system has been tested with inputs as low as 100 femtograms with no significant impact to the performance of the kit.
Q3: Why does the kit use only a single PCR amplification step?
This single PCR step combines targeted amplification of the microbial genome and the barcoding index PCR, which allows for a simple and fast workflow.
Q4: How does the kit achieve normalization?
The combination of using Equalase qPCR Premix to yield roughly equal amounts of libraries and a carefully curated barcode index list results in similar raw sequencing reads across samples.
Q5: What is the difference between the two standards included in the kit?
The ZymoBIOMICS Microbial Community DNA Standard consists of purified DNA from 8 bacteria and 2 yeasts and is useful for assessing community profile bias. The ZymoBIOMICS 16S/ITS qPCR Standard consists of a plasmid that has a single bacterial and single fungal target and is useful f http:// or quantifying copy number. Both are useful as positive controls for the library prep process.
Q6: Why is the library amplicon size not well defined in DNA fragment analysis?
Equalase amplification and the library prep design may cause some library products to not anneal well, causing a lack of a tight band. These libraries are perfectly fine because preparation for Illumina sequencing includes denaturing libraries into single strands.
