Illumina Genome Analyzer IIx
Dr. Alvaro Hernandez, Director of DNA Services
Our facility offers a full range of services for sequencing with the Illumina sequencing by synthesis technology. These include preparation of libraries from DNA or total RNA, sequencing and bioinformatic analysis.
The Illumina Genome Analyzer IIx utilizes an 8 lane flowcell and generates 6 to 20 million reads per lane with read-lengths of up to 76bp from one or both ends of the DNA fragments.
Applications for this technology include small RNAs, chromatin-immunoprecipitated DNA (ChIP_Seq), cDNA (mRNA_Seq), quantitation of gene expression (Digital Gene Expression, DGE), genome resequencing, comparative genetics and others.
For an overview of the technology please visit: http://www.illumina.com/pages.ilmn?ID=204
Sample submission requirements:
Sample Type |
Quantity |
Special instructions |
Genomic DNA |
1µg |
See gel images below for examples of high-quality DNA |
mRNA_Seq, DGE |
1-10µg of total RNA |
See gel images below. |
Chip_seq |
10ng |
|
Small RNA |
1-10ug of total RNA |
RNA should be extracted with Trizol, plant RNA reagent, mirVana, etc. RNA should not be purified with any column as this will eliminate small RNAs. |
Sequencing options:
- Single read or paired-ends: fragments from genomic DNA or cDNA are 200-500bp in length. These fragments can be sequenced from one end (single-read) or from both ends (paired-ends).
- Mate-pairs: sequencing fragments of genomic DNA that are 3kb to 5kb apart in the genome. This application is useful for assembly and orientation of contigs
- Barcoding: samples can be individually barcoded and multiplexed on a lane.
- 18bp to 76bp: the number of bases sequenced depends on the sample type, i.e. 18bp are sequenced from DGE, 36bp from small RNAs, whereas up to 76bp are sequenced from gDNA and cDNA.
Quality of DNA for gDNA library construction
High-quality gDNA on 1% Agarose Gel: 
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Low-quality gDNA on 1% Agarose Gel: |
Quality of RNA for cDNA, DGE or small RNA library construction
Example of high quality RNA. 28S rRNA is twice as intense as 18S rRNA.
Please contact Dr. Alvaro Hernandez, Director of DNA Services (aghernan@illinois.edu)
at 217-244-3480 or Chris Wright, Assistant Director of DNA Services (clwright@illinois.edu)
at 217-333-4372 to discuss ways the staff can be of assistance in achieving your project goals or to receive a quote for your project.
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High-Throughput Sequencing and Genotyping Unit
Director: Alvaro Hernandez, Ph.D.
340 Edward R. Madigan Laboratory, 1201 W. Gregory Drive, Urbana, IL 61801
Phone: (217) 244-3480 FAX: (217) 265-5066
Email: aghernan@illinois.edu
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| Last edited: 4 SEPT 2009 |
