Full-length cDNA library Production Service

• Product Description
• Price and Domestic Lead-time
• How to order
• Cautions

Strong tool for your transcriptome analysis such as gene expression profile analysis, function analysis and novel gene search.
This is a service to produce and deliver cDNA library with high full-length rate using the Vector-capping method (V-capping method *reference 1) with small amount of provided Total RNA.

This service is licensed for;

– Know-how license of National Rehabilitation Center for Persons with Disabilities and Hitachi High-Technologies Corporation

– Patent application, kokai number 2005-504229, by National Rehabilitation Center for Persons with Disabilities

Product Description

Using the “vector capping method”, cDNA library with high full-length rate (up to 95%) can be produced from small amount of Total RNA (ug order).

This service is introduced in the “nature” magazine (477-365-504 22 SEPTEMBER 2011).

– High full-length rate (95% at the maximum)

– Library production available from small amount of Total RNA (μg order)

– Library production available reflecting the expression level of each gene

– Highly effective to obtain long clones (*reference 2)

– The start point of transcription can be identified using the base G derived from cap structure.

Product Specification/Feature

– High full-length rate (95% at the maximum)

– Library production available with a small amount of Total RNA (ug order)

– Library can be produced with reflecting each gene’s expression level.

– Highly effective to obtain long chain clones (* reference 2)

– Transcriptional initiation point is discriminative. (using G derived from cap structure)

Once provided your purified total RNA sample, cDNA library will be made using the vector-capping method (V-capping method). When the library is made, 96 clones will be sequenced and the insert rate and full-length rate will be calculated. These are provided as evaluation data for your library purchase.

<Vector Capping Method>
This technology is developed by Dr. Seishi Kato of National Rehabilitation Center for Persons with Disabilities.

Reference

1. Kato, S., Ohtoko, K., Ohtake, H. and Kimura, T. 2005, Vector-Capping: A Simple Method for Preparing a High-Quality Full-Length cDNA Library. DNA Res., 12, 53-62.
2. Oshikawa, M., Sugai, Y., Usami, R., Ohtoko, K., Toyama, S. and Kato, S. 2008, Fine Expression Profiling of Full-length Transcripts using a Size-unbiased cDNA Library Prepared with the Vector-capping Method. DNA Res.15.123-136

< The Scope of the Service >

– QC of Total RNA. The result is to be reported.

– Production of full-length cDNA library

– Sequencing of 96 clones

– Full-length determination and insert sequence identification by BLAST search

– The identification result will be reported and the library solution will be delivered.

Service Flow

Sample to be provided

– Total RNA (extracted and purified: recommend to use RNeasy series from Qiagen)

– Concentration: more than 100ng/μL

– Total Amount:
For samples purified with RNeasy: more than 25μg
For samples purified in the other way: more than 40μg

– Shipping conditions:
1. Nuclease Free Water Solution (DEPC-treated water not acceptable), or,
2. Ethanol mixed solution (ethanol final concentration 80%; pl. do not add salt)

Deliverable

– Full-length cDNA library plasmid solution (dissolved in 50μL TE)

– 96 clones plasmid DNA (to be delivered in a 96 well plate)

– CD-R with the below contents;

1. cDNA library production report

2. 5′-end base sequence data (wave form of 96 clones, text data)

3. Result of Blast and ORF search (full-length determination excel data)

4. HTML file of each Blast search result

Price and Domestic Lead-time

Service Contents	Price	Domestic Lead-time
Full-length cDNA library production	Pl. inquire.	Please inquire.

How to order

Please inquire for details.

Cautions

– Description and scope of the service are subject to changes without notice.

– The fee of the work for evaluation data will be charged when no library purchase agreement is made after confirming the library evaluation data.

– Cancellation of an order will be rejected if received after production start. If cancellation is due to force majeure, the client will be asked to pay the service up to the cancellation point.

– Samples are limited to the RNA derived from class 1 and 2 nucleic acid donors defined in “the Law concerning the Conservation and Sustainable Use of Biological Diversity through Regulations on the Use of Living Modified Organisms” (Cartagena Protocol) issued by Ministry of Education, Culture, Sports, Science and Technology.

– In case of using human clinical sample, please confirm if

1. an approval through ethical review committee of the client’s affiliation is gained.

2. an informed consent (consent from the sample donor) is obtained.

3. Anonymization (data is not linked to personal information through electric data etc.) is made.

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