Up to 10 000 PCR reactions/week, all controlled on agarose gel

We offer the following services: 

1-Pair of primer design targeting specific region of genes 

2-PCR amplification of cDNA inserts and amplicons using DNA genomic as templates 

3-DOP-PCR productions on BACs (Bacteria artificial chromosome) for CGH Array

 4-All the PCR reactions are controlled on agarose gel for specificity and yield 

5-Automatisation of PCR purification, aliquoting and quantification 

6-Spotting on either glass slides, nylon membranes or 96 well plates 

7- Microarrays-quality control

8- -Hybridization services and data analysis

Name	Quantity	Reference	EUR
PCR amplification services	1	AR-PCR-01	Upon request

Pair of primer design 
Eurogentec offers primer design service for customized PCR MicroArrays. The design of the PCR primers is performed on the complete genome sequence to amplify unique region for every coding sequence of the genome or for only a particular subset of genes of your interest. The customer can provide a list of Gene Bank accession information or the sequence information in a Fasta and txt format. Process of primer design Each entire ORF will be analyzed for sequence similarity using a Blastn search against a database containing the genome sequence information. The orthologus genes will be analyzed carefully in order to identify a unique region, which will be represented by shorter amplicon. 
Typical Design Criteria 
• All primer pairs for different ORFs will have similar Tm, i.e., 56 ± 5 °C. This greatly facilitates amplification of a large number of fragments, as the PCR conditions are homogeneous across ORFs. Amplicons generated using the selected primers will have similar sizes within 150 to 700 bp. This allows consistent yields among different amplicons. 
• Candidate primers undergo a test against the rest of the genome and are redesigned if high complementarily secondary site(s) is (are) found. • We automatically run Blastn search of all the candidate amplicons against all the sequences of ORFs. Typically, the amplicons are redesigned if they have more than 20 % sequence identity with other ORFs. PCR reactions We offer either single or two-step method of PCR reaction to amplify specific region of ORFs. We use a two-step method of PCR reaction as shown in figure 1: 
• Primary PCR: The PCR reactions are performed using tagged genome specific primers in the presence of genomic DNA (provided by your laboratory). We normally achieve a success rate of 93 % of PCR reactions. For the failed PCR reactions we perform three different protocols of PCR conditions, which include different MgCl2 concentrations and different Taq polymerase. We should normally get an additional 2 % working and reach a success rate of 95 % in total. We check 5 % of the total oligos by MS and will re-synthesize maximum 2 % of the oligos based on a newly designed primer. We will perform the PCR reactions with the new designed oligonucleotides with the standard PCR protocol. Following this process we reach usually 97 % success rate. 
• Secondary PCR: amplifications will be performed using a single pair of primers annealing to tag sequences introduced on primary PCR primers. These PCR amplifications will also be controlled by agarose gel electrophoresis and repeated if required reaching a minimum of 95 % success rate. The final PCR products will contain an amino group at their 5’ end to promote the attachment of the predicted gene coding DNA strand to the treated glass slides.

• For specific quote and more information, please contact Dr Driss Talibi:
  
  dr.talibi@eurogentec.com
  Tel.: +32 4 372 74 39
  Mobile: +32 478 366 619
  Fax: +32 4 372 75 00

• Online
  You can place your order online. Login to access our web ordering interface.

• By e-mail: order@eurogentec.com

• By fax : +32 4 264 07 88

• By mail
  Eurogentec S.A.
  LIEGE Science Park
  Rue du Bois Saint Jean 5
  4102 Seraing
  BELGIUM

If you still have questions with ordering please contact our Customer Support : +32 4 372 76 65.