Index of /public/revio/2025Q2/microbes/metagenomics
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README.txt 2025-06-17 15:32 1.9K
SAMPLES
SAMPLE PROTOCOL BARCODE PAIR NAME
------ -------- -------
Fecal reference (Zymo D6323) WGS bc2001
Fecal reference (Zymo D6323) Ampli-Fi Plate_A_1_A01
Gut microbiome standard (Zymo D6331) WGS bc2002
Gut microbiome standard (Zymo D6331) Ampli-Fi Plate_A_2_B01
METAGENOMIC SAMPLE POOL PREPARED WITH AMPLI-FI PROTOCOL AND WGS PROTOCOL
METHODS
4 libraries were prepared: fecal reference (Zymo D6323) and gut microbiome standard (Zymo D6331) using both the Ampli-Fi protocol and WGS protocol for each sample
PROTOCOL Amplifying genomic DNA for SMRTbell library preparation and HiFi sequencing (103-648-000 REV2 MAR2025)
Preparing whole genome and metagenome libraries using SMRTbell prep kit 3.0 (102-166-600 REV06 JAN2025)
INPUT Ampli-Fi protocol: 45 ng/sample
WGS protocol: 2 ug/sample
SHEARING Not applicable: fecal reference stock sized at 10.1 kb and gut microbiome standard stock sized at 7.4 kb
BARCODING Ampli-Fi protocol: Twist Universal Adapter System TruSeq Compatible, 96 Samples Plate A (101308)
WGS protocol: SMRTbell adapter index plate 96A (102-009-200)
SEQUENCING Vega system, 200 pM loading concentration, 24 hr movie (m21015_250606_023823)
Revio system, 225 pM loading concentration, 24 hr movie (m84039_250606_030410_s4)
ANALYSIS WGS Protocol:
Demultiplex SMRTbell adapters with lima v2.12.0: `lima --split-bam-named --preset SYMMETRIC-ADAPTERS`
Ampli-Fi Protocol:
Demultiplex Twist UDI adapters with lima v2.12.0: `lima --split-bam-named --preset ASYMMETRIC`
Remove PCR duplicates with pbmarkdup v1.1.0: `pbmarkdup --hifi --remove-pcr-duplicates`