| LightCycler® Application Service |
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| Optimization of LightCycler® |
It is said that thermal cycling of LightCycler® is the fastest in the world.
Temperature control and homogeneity of optical detection of LightCycler®480 is very superior.
We will provide you with high-quality applications using LightCycler® and LightCycler®480 based on accumulated technology of real-time PCR for many years.
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| Compliant Applications |
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| Purpose | Method |
| Quatitation of Gene Expression (mRNA Quantitation) | 1step qRT-PCR |
| 2step qRT-PCR |
| DNA Quantitation | qPCR |
CNV Analysis
Gene amplification
Quantitation of virus · bacteria | qPCR |
| SNP Analysis | Melting Curve Analysis by HybProbe Method |
| HRM Method |
| Quantitation of Mutated DNA | Please consult us. |
| Quantitation specific to Alternative Splicing Variant | Please consult us. |
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| * We can accept other purposes that the above list. |
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| Biological Species |
| We can deal with all kinds of biological species. |
| Example) human, mouse, rat, bovine, swine, chicken, zebrafish, drosophila, Arabidopsis, rice, fungus, bacteria, virus & others. |
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| Detection Method |
| You can select either method from the followings. |
- Intercalator Method (SYBRGreen I)
- HybProbe Method
- Hydrolisis Method
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| Three Characteristics of HybProbe Method |
- High specificity due to detection by 2 kinds of probes.
It is necessary that two probes need to hybridize to amplified products in order to obtain fluorescence, so it can be said that it has high reliability than 1 kind of probe.
- Long Amplicon-size can be used, too.
There is no limitation in Amplicon size and base at 5’ terminal as hydrolyzed probes.
It is usable when Amplicon size cannot be set due to Alternative Splicing Variant and homologous genes.
- Possible to undergo polymorphism analysis and detection of mutation by melting curve analysis.
When there is polymorphism and mutation in the probe region, melting temperature between amplified products will shift to the side of low temperature, so it becomes possible to undergo analysis of polymorphism and detection of mutation by performing melting curve analysis.
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| Principle of HybProbe Method |
As a result of hybridization between probes labeled with LCRed at 5' terminal and other probes labeled with fluorescein at 3’ terminal, the two probes come close and fluorescence resonance energy transfer (FRET) will occur.
Donor dye fluorescein is excited by light source of LightCycler® and a part of excited energy is transferred to acceptor dye, LCRed, then fluorescence will be emitted from it. Fluorescence of LCRed is measured by LightCycler®. |
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| Price / Delivery Time |
Following is the list of price and delivery time of standard applications.
We accept it as Basic Course.
We accept applications other than the followings as separate course and will quote the price and delivery time depending on the content of request.
| Application | Price | Delivery Time (For Japan) |
| mRNA Quantitation(2step, use SYBRGreen I) | ¥60,000 | 2 weeks after receipt of samples |
| mRNA Quantitation(2step, use HybProbe) | ¥84,000 | 3 weeks after receipt of samples |
| mRNA Quantitation(2step, use Hydrolysis Probe) | ¥84,000 | 3 weeks after receipt of samples |
| DNA Quantitation(2step, use SYBRGreen I) | ¥50,000 | 2 weeks after receipt of samples |
| DNA Quantitation(2step, use HybProbe) | ¥74,000 | 3 weeks after receipt of samples |
| DNA Quantitation(2step, use Hydrolysis Probe) | ¥74,000 | 3 weeks after receipt of samples |
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| * Prices are excluding tax. Consumption tax is put on the price separately. |
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| What to prepare: |
| It's necessary for you to send positive samples for examining conditions. |
| Application | Contents of Mailing |
| mRNA Quantitation(2step, use SYBRGreen I) | Total RNA 30µg |
| mRNA Quantitation(2step, use HybProbe) |
| mRNA Quantitation(2step, use Hydrolysis Probe) |
| DNA Quantitation(2step, use SYBRGreen I) | Genomic DNA (10ng/µL)30µL |
| DNA Quantitation(2step, use HybProbe) |
| DNA Quantitation(2step, use Hydrolysis Probe) |
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| We will E-mail you details how to send samples after receipt of your application. |
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| Optimization of LightCycler®480 Dual-Plex |
It is possible to detect many fluorescent dyes by LightCycler®480, so it's very suitable for multiplex PCR assay.
Based on our accumulated knowledge of PCR, we can offer quantitative analysis of genes as well as SNP analysis.
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Depending on your request,
We will provide you with most optimal detection method (HybProbe Method or Hydrolyzed Probe Method ).
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| Compliant Applications |
| Purpose | Method |
| Quantitiation of Gene Expression( mRNA Quantitation | 1step qRT-PCR |
| 2step qRT-PCR |
| DNA Quantitation | qPCR |
CNV Analysis
Gene Amplification
Quantitation of Virus · Microorganisms | qPCR |
| SNP Analysis | Melting Curve Analysis by HybProbe |
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| representative fluorescent dye |
| Reporter / Quencher | Ex / Em |
| 5'-6FAM / 3'-ATTO540Q | 494 / 525 |
| 5'-6HEX / 3'-ATTO540Q | 535 / 556 |
| 5'-6HEX / 3'-BHQ-1® | 535 / 556 |
| 5'Yakima Yellow / 3'-BHQ-1® | 530 / 549 |
| 5'-LightCycler®480Cyan500 / 3'-BHQ-1® | 450 / 500 |
| 5'-6FAM / 3'-BHQ-1® | 483 / 533 |
| 5'-LightCycler®Yellow555 / 3'-BHQ-1® | 535 / 568 |
| 5'-LightCycler®Red610 / 3'-BHQ-2® | 558 / 610 |
| 5'-LightCycler®Red670 / 3'-BHQ-2® | 615 / 670 |
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| Price / Delivery Time |
| We will quote depending on desired applications. |
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| What to prepare: |
| It's necessary for you to send positive samples for examining conditions. |
| Application | Contents of Mailing |
| mRNA Quantitation | Total RNA 30μg |
| DNA Quantitation | Genomic DNA (10ng/μL)30μL |
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| We will E-mail you details how to send samples after receipt of your application. |
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| Design and Synthesis |
We can deal with all kinds of applications for real-time PCR.
We will provide you with high-quality designs of primers and probes based on our unique design standards.
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| Compliant Applications |
- Quantitation of Gene Expression (mRNA Quantitation)
- Quantitation of Gene Expression (effect of siRNA confirmed)
- Quantitation of Gene Expression (effect of microarray confirmed)
- DNA Quantitation
- CNV Analysis
- Gene Amplification
- Auantitation of Virus · microorganisms
- SNP Analysis (HRM Method)
- SNP Analysis (HybProbe method)
- Quantitation of mutated DNA
- Quantitation specific to alternative splicing Variant.
- ChIP qPCR
- Designing of Primers for Sequence Analysis
* We are happy to comply with other inquiries.
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| Price / Delivery Time |
| Delivery Time: Within 5 working days for japan. For overseas, please contact us |
| * It is a delivery time for orders within 10 genes in number of genes. |
| If crowded with many orders, it might take more than that time. |
| In case of urgency, please inquire at the time of application. |
| Price: Design Price is free of charge. |
Synthesis: Please see price list for synthesis.  Price list for synthesis |
| * It costs you design fee if you would like to use instruments other than LightCycler® |
| t casts you sequence analysis fee in case you need specific sequence analysis. |
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| What to prepare: |
| GenBank Access No. of target genes |
| (rs No. in case of SNP) |
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| Preparation of Standard( DNA/RNA Standard) |
We provide you with Standard DNA and RNA corresponding to applications of real-time PCR.
It is the absolute condition for standards that they have to be stable.
Preservation stability of RNA standard has improved dramatically by the use of stabilizer uniquely developed by us ( patent applied).
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| Kinds of standards and applications |
| Appliation | Standard |
| 2step RT-qPCR | Plasmid DNA |
| 1step RT-qPCR | artificial RNA(in vitro transcripion) |
| DNA Quantitation | Plasmid DNA |
| SNP Analysis | PlasmidDNA |
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| Price / Delivery Time |
| Standard | Price | Delivery Time (For Japan) |
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| Plasmid DNA | ¥650,000 – | 1 Week – |
| Artificial RNA | ¥ 76,000 – | 2 Weeks – |
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| Whats to prepare |
| It's necessary for you to send positive samples. |
| Application | Contents of Mailing |
| mRNA Quantitation | cDNA Over20µL
(reverse transcription product of the corresponding totalRNA 1µg) |
| DNA Quantitation | Genomic DNA (10ng/µL)30µL |
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| We will E-mail you details how to send samples after receipt of your application. |
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| Contact Information |
Contact Information |
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