New automatic DNA sequencing and gene characterisation (reading of fragments) service.

Test equipment
In order to keep constantly abreast of the times, the Istituto Nazionale di Apicoltura has equipped its laboratories with instruments used in molecular biology.
The main instrument of analysis is a modern automatic sequencer (CEQ8000 Beckman Coulter), which enables us to analyse DNA sequences and DNA fragments (microsatellites, SNPs, AFLPs).

Shipping instructions

Sequences
Reading only
After the sequencing reaction, ethanol precipitate the reaction or purify with the special kits recommended by the manufacturer (Beckman Coulter). Fill in the sequence – read only order form and send it together with the test tubes, which should be sealed with parafilm and wrapped with foil (the pellet is photosensitive!!). If the samples are to be dispatched already resuspended in SLS, pack the test tubes on wet ice and make sure the samples are sent as promptly as possible (within 48 hours).

Sequencing reaction and reading
Purify the PCR product by enzymatic reaction (Exo-SAP) or using one of the commercially available kits designed for this purpose; quantify the concentration of the PCR product by agarose gel electrophoresis and/or spectrophotometry. Send a sufficient quantity for at least two reactions (2X). The quantity of template to be sent will depend on the type of DNA (plasmid dsDNA, M13 ssDNA, PCR products...): as a rule, for dsDNA and purified PCR products you should refer to the table under the heading “ng per 100 fmol”; for ssDNA refer to the table under the heading “ng per 50 fmol”. Send the primers at a concentration of 1.6µM, in adequate quantity (2.0µl is required for each reaction). Fill in the sequence – reaction + reading order form and send it together with the test tubes, which should be sealed with parafilm and placed on wet ice, within a few days.

Fragments (reading only)
Send at least 10 µl of PCR product (microsatellites, SNPs, AFLP) marked with the dye-primer. If you wish to have more than 4 fragments run simultaneously, purify the PCR product using one of the commercially available kits designed for this purpose or an ethanol/sodium acetate precipitation. Fill in the fragments order form and send it together with the test tubes, which should be sealed with parafilm, wrapped with foil and placed on wet ice, within a few days.
If you wish the laboratory to perform the amplification reaction as well, please contact the technician before sending the samples.

> Prices of analyses (click here)

NOTE: should you have any doubts or require any explanation, or wish to discuss special projects and needs, please do not hesitate to contact the technician (Raffaele Dall’Olio)