The EQUAL-seq Programme

1. Objectives of the EQUAL-seq Programme

To-date, external quality control programmes (EQA) designed to monitor PCR and PCR-related assays have been very limited. EQAs, especially designed for sequencing based methods have not been undertaken until yet.

Within the EQUAL-seq programme we intend to provide three kinds of samples to be investigated by the participant:

2. Participant selection

Participation is open to all laboratories carrying out sequencing based methods within the EU. These can be Clinical Laboratories of different medical professions and Industry Laboratories.

Please note, that there is a restriction of 50 participants.


3. What do you need to participate?

Sequencing

You will be requested to use your DNA sequencer (genetic analyzer) and the reagents commonly used in your laboratory.

Uploading of your data

You will be requested to insert your data in the DGKL web-site (www.DGKL.de). You will go through a short questionnaire concerning the methods used. Text strings and raw data files will be necessary to report your results.

Send an aliquot of sample 1 and 4

Purified products 1 and 4 must be sent back to the coordinator!

Please send the aliquots back by express mail to the EQUAL-seq project management (Shipment box enclosed in the kit) for further analyses. Please note that the expenses for this expedition must be covered by your institution.

Downloading of the results

Your performance and those of all participants will be evaluated anonymously and will be consultable on the DGKL web-site (www.DGKL.de). Additionally, you will receive comments by the organizer.


4. What will you find in the provided EQUAL-seq kit? Which actions do you have to do?

This is the list of the material included in the EQUAL-seq kit and of the actions to be performed in your laboratory.

Sample/
reagent
 quantity Volume / concentration Actions
     
Sequen-
cing
Text file
Abi
Traces
Note/ comment

1. Plasmid DNA

 1 + primer to use in sequencing reaction 2 µg(Plasmid DNA in H2O)
X
X
X
X

2. Plasmid DNA

 1 + primer to use in sequencing reaction 2 µg (Plasmid DNA in H2O)
X
X
X
X

3. PCR-Product

 1+ primer to use in sequencing reaction 300ng
X
X
X
X

4. sequencing reaction product

 1 one sequencing reaction
X
X
X
X
Total actions    
4
4
4
4


5. Description of the samples and the actions

Sample #1 will be plasmid DNA. The participant is asked to perform the sequencing reaction and to provide the longest possible sequence without any mistakes (e.g. “N”). The primer will be provided by the coordinator.
The analysed reaction of this sample is to be sent back to the coordinator for further investigation.

Sample #2 will be plasmid-DNA. The participant is asked to perform the sequencing reaction and to provide the longest possible sequence without any mistakes (e.g. “N”). The primer will be provided by the coordinator.

Sample #3 will be a PCR-Product. A primer will be sent out to perform the reaction as well.

Sample #4 will be a completed sequencing reaction which should be purified and analysed by the participant. We will provide a finished reaction established in our laboratory. In this case, of course, no sequencing primer will be provided. The analysed reaction of this sample is to be sent back to the coordinator for further investigation.

An indispensable prerequisite for these experiments is that purified reactions from samples 1 and 4 are sent back to the coordinator to evaluate differences in technical equipment and methods!


6. Time Schedule of the Survey

Deadline or application November, 6, 2004
Shipment of the EQUAL-seq kit to participants starts October 2004
ends December 2004
Phase of analysis in the labs, uploading of the results and shipment of the samples October, 2004 - February, 2005
Closing of the survey February 26, 2005
Re-evaluation of materials at the facilities of the organizers until end of April 2005
Statistical analysis on the Web site May 2005
Back-Reporting May 2005
Training Courses planned in the period July - October 2005

7. Do you want to know more on it?

Are there any questions? Do you need further informations? Please send a message to Prof. Dr. med. Michael Neumaier by e-mail (michael.neumaier@ikc.ma.uni-heidelberg.de) or by mail to:

Prof. Dr. med. Michael Neumaier
Institute for Clinical Chemistry
Faculty of Clinical Medicine Mannheim, University of Heidelberg
Theodor-Kutzer-Ufer 1-3
68135 Mannheim
Germany

8. Do you want to join the EQUAL-seq programme?

Please fill your application form by clicking here.