How does the Illumina MiSeq work?

How does the Illumina MiSeq work?

Sequencing is performed by recording the synthesis of DNA strands in clusters of sample templates attached to the flow cell. Each newly attached base liberates a fluorescent dye that is excited by diode lasers (530 & 660 nm) and imaged using two digital cameras.

What is Illumina sample sheet?

The Sample Sheet is a file format used by Illumina for storing biological sample information and metadata associated with a given experiment. This file is used widely across the Illumina informatics ecosystem as an input to many pieces of software, such as bcl2fastq and BaseSpace® Sequence Hub.

How do you power cycle a MiSeq?

To power cycle in Windows mode:

  1. Close MiSeq Control Software and ensure no other programs are running and shut-down the computer from the Windows Start button.
  2. Once the computer has shut down, turn off the power switch on the back of the MiSeq instrument, and leave it off for a minimum of 60 seconds.

What is sample sheet?

The sample sheet is a comma-delimited file (SampleSheet. csv) that stores the information needed to set up and analyze a sequencing experiment. The file includes a list of samples, their index sequences, and the sequencing workflow.

What is the purpose of Illumina sequencing?

Sequencing may be utilized to determine the order of nucleotides in small targeted genomic regions or entire genomes. Illumina sequencing enables a wide variety of applications, allowing researchers to ask virtually any question related to the genome, transcriptome, or epigenome of any organism.

What is Illumina Experiment Manager?

The Illumina® Experiment Manager (IEM) is an app used to create valid sample sheets for some Illumina® sequencers. Although sample sheets can be generated manually, this tool minimize the risk of errors. By default, only Illumina® indices are preprogrammed in the software.

What should I do if the MiSeq fails to measure flow rate when starting a run?

If the flow check still fails, power cycle the instrument:

  1. Close MiSeq Control Software, shut down Windows, and turn off the instrument using the power switch on the back panel.
  2. Leave the instrument off for at least 2 minutes, to allow all parts to completely power down.

What is Illumina BaseSpace?

BaseSpace is a powerful website where biologists and informaticians can easily store, analyze, and share genetic data. Sequencing data from Illumina instruments, including MiSeq and HiSeq, are streamed in real-time over the Internet to BaseSpace.

What are the applications of Illumina sequencing?

How to prepare a library for MiSeq sequencing?

Prepare your libraries as described in the protocol for TruSeq Exome Enrichment Kit or the TruSeq Custom Enrichment Kit, and prepare your libraries for sequencing as described in the MiSeq System Guide. When you are ready to set up your sample sheet using the Illumina Experiment Manager, select the following settings:

How are reagents loaded into the MiSeq system?

All reagents for cluster generation, sequencing, and paired-end chemistry are loaded onto the instrument in a pre-filled reagent cartridge prior to starting the run. When the run is started, the MiSeq System performs cluster generation followed by sequencing and paired-end chemistry (if applicable).

How much template is needed for a MiSeq system?

The MiSeq System uses a total of 500 µl of denatured and diluted template for priming and clustering. However, 600 µl of template is recommended to avoid air bubbles and accommodate for instrument sipper depth. This volume does not vary for different workflows.

What are instructions for operating MiSeq in Windows 7?

System overview and instructions for operating and maintaining the MiSeq System on Windows 7. System overview and instructions for operating and maintaining the MiSeq System. Instructions for preparing PhiX and denaturing and diluting libraries for sequencing on the MiSeq.