Dna Sequencing Service

Q: What type of Sequencing Service does Primm offer?

A: Basic Read: Customer provides DNA and oligos premixed. Sequencing results are supplied as complete electropherograms checked for the absence of technical or elecrophoretic problems. Customer performs data editing.
Smart/Quick Read: Customer provides DNA and primers in a 1,5 ml tube in a maximum amount of 10 ul. Sequencing results are provided as complete electropherograms checked for the absence of technical or elecrophoretic problems. Customer performs data editing.
Safe Read: Customer provides DNA and primers in a 1,5 ml tube in a maximum amount of 10 ul . Sequencing results are supplied as complete electropherograms together with a text file. Primm performs the editing of all sequencing results.
HT Read: High-throughput-96-well-format plates sequencing.
Walking Primer Strategy: Primm provides sequencing results as complete electropherograms together with a text file and a contig of the all performed sequences.
Loading Sample: Customer provides performed sequencing reaction. All the reaction should be purified, lyophilized or dissolved in a maximum amount of 10 . Primm can process only sequencing reactions performed with Applied Biosystem Big Dye Chemistry v3.1.

Q: Where can I send my DNA samples?

A: All DNA samples can be sent to:
PRIMM srl
Via Olgettina, 58
20132 Milano

For the customer based in Milan a pick-up service is available.

Q: How much DNA should I submit?

A:

For any further information please consult our technical detailed lists.

Q: How should I submit DNA for sequencing ?

A: Please submit samples in 1,5ml tube. It is not necessary to send the samples on ice and use ordinary mail or other carrier service.

Q: How should I label my tubes?

A: Please label all the tubes with the ID number together with sample’s or primer’s name.

Q: What is the best way to determine DNA samples concentration?

A: One of the most crucial factor for successful sequencing reaction is to use the right amount of template. For this reason we suggest to check the concentration of your samples loading them onto an agarose gel rather then measuring UV absorbance.

Q: How should I prepare and purify my plasmidic template for sequencing?

A: It’s extremely important to send samples pure and free from contaminants (salts, ethanol, etc..).In order to obtain results of good quality, we suggest to prepare your templates according to standardized methods using commercial kits suitable for DNA sequencing analisys. Use always ddH2O or Tris-HCl 10mM to resuspend your DNA.

Q: Can I send unpurified PCR products?

A: PCR fragments must be single band. Our service performs a free ExoSAP-IT® treatment on every PCR product before setting up the sequencing reaction (not available on “Basic Read” service). If aspecific bands are present, please purify your samples using commercial kits suitable for DNA sequencing analisys before submit them to our service.

Q: Can I send bacteria?

A: Customer can supply bacteria as:
• O.N. culture in LB or like medium, minimum amount 5 ml for each single reaction (50 μl for HT-Read)
• Single colony, on a petri plate or a tube stub.
It is mandatory to indicate the antibiotic resistance of the E. Coli strain.
Template preparation from Bacteria is performed using a commercial kit suitable for sequencing reaction set-up.

Q: What primers does Primm provide?

A: A list of the most common universal primers is available on the website. The use of these primers is free of charge.

Q: How should I submit my custom primers?

A: Specific oligonucleotides can be ordered using our web site or provide by the customers.
Provided oligos can be dissolved in ddH2O at the concentration of (10 for each sequencing reaction).

Q: Which features should my sequencing primer have?

A: Primers that give best results in automated sequencing must have a Tm between 50°-55°C and should not be rich in C or G at the 3' end but can contain C or G as the final 3' base.

Q: When should I order Walking Primer service?

A: We suggest to use the “Walking Primer Strategy” to sequence fragments up to 3Kb. This type of sequencing service should be used for PCR fragments or plasmids.

Q: How long will my sequencing data be stored?

A: Your data will be stored for one year. To prevent loss of data we suggest to download your sequencing results on your PC.

Q: How long does Primm store samples and primers?

A: The DNA and the primer should be stored for two months.

Q: How can I order Primm sequencing service?

A: Use our web site to order sequencing reaction. It is mandatory to register your data before using the ordering online system -POLOS-.

Q: How can I order plate sequencing?

A: It is possible to order sequencing reactions in 96 well format plate (HT-Read sequencing service). This service is available for at least 48 samples/plate.

Q: Where can I obtain the programs to visualize the chromatograms?

A: Different freeware software to visualize and analyse your sequencing results are available on our website:

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