Encore® BiotinIL Module
The Encore® BiotinIL Module is designed to provide a rapid, simple and easily automatable approach for labeling amplified cDNA generated with NuGEN® amplification systems for analysis on the Illumina Genome-Wide Expression BeadChips.
- Label amplified cDNA in about 2 hours for hybridization onto Illumina Genome-Wide Expression BeadChips
- Simple add and incubate procedure requires only one purification step
- Compatible with amplified cDNA generated with NuGEN’s Ovation® and Applause® amplification systems
The Encore® BiotinIL Module provides optimized reagent mixes and protocol to process 48 cDNA samples.
High sensitivity and labeling efficiency
- High reproducibility and call concordance
- Gene expression
|Compatible Platform||Illumina BeadChips|
|Amplification Type||Not applicable|
|Starting Material||Amplified cDNA|
|Input Amounts||750ng – 1.5ug|
The Module provides all necessary buffers and enzymes for labeling of cDNA generated with a compatible NuGEN RNA Amplification System.
What equipment is required or will be useful?
Required equipment includes a microcentrifuge, pipettes, vorteer, a thermal cycler, and a Nanodrop or UV/Vis spectrophotometer. An Agilent Bioanalyzer or a similar instrument may be used for quality control.
What additional reagents are required for the Encore BiotinIL Module?
No additional reagents are required, but MinElute purification columns are required for the final purification step (see Appendi A of the User Guide).
Has NuGEN performed reproducibility studies on the Encore BiotinIL Module?
Yes, our studies have demonstrated high sample-to-sample, lot-to-lot and operator-to-operator reproducibility.
You must use SPIA cDNA generated with one of the following NuGEN products with the Encore BiotinIL Module:
- Ovation RNA Amplification System V2 (Part No. 3100)
- Ovation Whole Blood Solution (Part Nos. 1300 and 3100)
- Ovation Pico WTA System V2 (Part No. 3302)
- Ovation PicoSL WTA System V2 (Part No. 3312)
- Ovation FFPE WTA System (Part No. 3403)
- Ovation One-Direct System (Part No. 3500)
- Applause WT-Amp Plus ST System (Part No. 5510)
Can I vary the amount of cDNA input for labeling?
The recommended cDNA input amounts range from 2 to 4 μg.
Can I use any cDNA as starting material in the Encore BiotinIL Module?
No, the cDNA must be generated using a compatible NuGEN RNA Amplification System. Use of other cDNAs will result in poor performance.
Can the Encore BiotinIL Module be used for labeling of RNA?
No. The Encore BiotinIL Module is intended to label amplified cDNA, not RNA. RNA is not compatible with the labeling chemistry employed by this Module.
We do not recommend stopping at any step of the fragmentation or labeling protocols.
What are the recommended storage conditions for the labeled cDNA?
The labeled cDNA should be stored at –20°C. Ensure the vials are well sealed and avoid multiple freeze/thaw cycles.
You should epect to recover greater than 80% of your input cDNA following purification.
How should I qualify input cDNA for use with the Encore BiotinIL Module?
The concentration of starting cDNA must be determined to ensure adequate input into the labeling reaction and, therefore, onto the arrays (see Appendi D of the product User Guide). You may choose to further qualify the starting cDNA by performing qPCR assays as recommended in the appropriate NuGEN Amplification System Technical Reports.
The Encore BiotinIL Module has been validated for use only on Illumina Whole Genome Epression BeadChips.
Are the array hybridization reagents included in the Encore BiotinIL Module?
No. We only provide the reagents necessary for labeling of the cDNA. We do provide a recommended procedure for hybridization. See Appendi C of the User Guide.
What hybridization and wash protocols do you recommend for Illumina Whole Genome Epression BeadChip applications?
We recommend following the array manufacturer’s protocol with the following adjustments: a. Hybridize arrays at 48°C instead of the manufacturer’s recommended temperature of 58°C.
How much labeled cDNA should I hybridize to an Illumina Whole Genome Epression BeadChip array?
We recommend hybridizing 750 ng to 1.5 μg labeled cDNA. See Appendi C of the User Guide.
Should I purify the cDNA before hybridization?
Yes. It is necessary to purify the labeled cDNA prior to hybridization to minimize background signal on the array (See Appendi A of the User Guide).
For research use only. Not for use in diagnostic procedures.