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Illumina pre-made libraries

Illumina sequencing technology, based on Sequencing by Synthesis (SBS), is a globally embraced NGS innovation, responsible for generating over 90% of the world’s sequencing data. The principle of SBS involves imaging fluorescently labelled reversible terminators as each dNTP is added, and subsequently cleaved to allow the incorporation of the next base. With all four reversible terminator-bound dNTPs present in each sequencing cycle, natural competition minimizes incorporation bias. This versatile technology supports both single-read and paired-end libraries, catering to a range of genomic applications. Illumina sequencing’s high-throughput capabilities and precision position it as a cornerstone in genomics research, empowering scientists to unravel the intricacies of genomes with unmatched detail and efficiency.

Our pre-made library sequencing service enables customers to prepare sequencing libraries from diverse sources (mRNA, whole genome, amplicon, 10x libraries, among others). Subsequently, these libraries can be shipped to our sequencing centers for quality control and sequencing in Illumina platforms.

Service Details

Demo Result

Features

● Platforms: Illumina NovaSeq 6000 and NovaSeq X Plus

● Sequencing modes: PE150 and PE250

● Quality control of libraries before sequencing

● Sequencing data QC and delivery: delivery of QC report and raw data in fastq format after demultiplexing and filtering Q30 reads

Service Advantages

● Versatility of Sequencing services: the customer may choose to sequence by lane, flow cell, or by amount of data required (partial lane sequencing).

● Extensive experience on Illumina sequencing platform: with thousands of closed projects with various species.

● Delivery of sequencing QC report: with quality metrics, data accuracy and overall performance of the sequencing project.

● Mature sequencing process: with short turn-around time.

● Rigorous Quality Control: we implement strict QC requirements to guarantee the delivery of consistently high-quality results.

Sample Platforms

Platform	Flow Cell	Sequencing mode	Unit	Estimated output
NovaSeq X	10B (8 lanes)	PE150	Single Lane Partial Lane	375Gb / Lane
NovaSeq X	25B ( 8 lanes)	PE150	Single Lane Partial Lane	1000 Gb/Lane
NovaSeq 6000	SP Flow cell (2 lanes)	PE250	Flow Cell Single Lane Partial Lane	325-400 M reads / Lane
NovaSeq 6000	S4 Flow cell (4 lanes)	PE150	Flow Cell Single Lane Partial Lane	~800 Gb / Lane

Sample Requirements

	Data Amount (X)	Concentration (qPCR/nM)	Volume
Partial Lane Sequencing	X ≤ 10 Gb	≥ 1 nM	≥ 25 μl
	10 Gb < X ≤ 50 Gb	≥ 2 nM	≥ 25 μl
	50 Gb < X ≤ 100 Gb	≥ 3 nM	≥ 25 μl
	X > 100 Gb	≥ 4 nM	≥ 25 μl
Lane Sequencing	Per Lane	≥ 1.5 nM / Library pool	≥ 25 μl / Library pool

In addition to concentration and total amount, a suitable peak pattern is also required.

Note: Lane sequencing of low diversity libraries requires PhiX spike-in to ensure robust base calling.

We recommend submitting pre-pooled libraries as samples. If you require BMKGENE to perform library pooling, please refer to

the library requirements for partial lane sequencing.

Library Size (Peak map)

Main peak should be within 300-450 bp.
Libraries should have a single main peak, no adapter contamination and no primer dimers.

Please reach out to us if your samples don't meet the starting material requirements.

Service Workflow

Library quality control

Sequencing

Data quality control

Project delivery

Previous: DNA/RNA sequencing -PacBio Sequencer

Next: DNA/RNA Sequencing – Nanopore Sequencer

Library QC report

A report on the quality of the library is provided before sequencing, assessing library amount, and fragmentation.

Sequencing QC report

Table 1. Statistics on sequencing data.

Sample ID	BMKID	Raw reads	Raw Data (bp)	Clean reads (%)	Q20(%)	Q30(%)	GC(%)
C_01	BMK_01	22,870,120	6,861,036,000	96.48	99.14	94.85	36.67
C_02	BMK_02	14,717,867	4,415,360,100	96.00	98.95	93.89	37.08