Nextera XT low complexity pooling, V3 reagent kit

[lac302]

A couple questions about the Nextera XT kit. The protocol is written as if you are using amplicons.

1. What is the size distribution of fragments post tagmentation assuming loading 1ng of gDNA (metagenome)?

2. What is the binding capacity of the normalization beads?

3. If pooling 1-3 libraries how much of each should be added the HT1 buffer. I was told by tech support to load only 5ul of ea library, bring up the volume of HT1 to 600ul. This doesn't seem right to me. Protocol states 5ul of ea library pooled, 24ul of this into 576 HT1 to maintain 1:25 dilution.

[nucacidhunter]

1- Tagmented fragments size using XT kit for good quality high molecular weight DNA is 100-2000 bp. Completed libraries sizes varies from 300 bp above depending on ratio of beads used for clean-up.
2- Illumina does not reveal that.
3- We stop after cleaning PCR and follow standard protocols for sequencing which is QPCR quantification, pooling and sequencing. Using normaliser beads gives unreliable results.