1. DNA preparation

Pure DNA is essential for good sequences.

Please use commercially available kits to prepare your plasmid-, phage-, or BAC/PAC-DNA. The DNA should be ethanol-precipitated at room temperature after the last preparation step. Dissolve the pellet in water or EDTA-free buffer.

All PCR-products must be purified. If the product is homogeneous it is usually sufficient to remove the amplification primers by PCR purification kits. Contaminating by-products have to be removed from non-homogeneous PCR products by agarose gel purification.

Both purification methods can be done by us, if requested.

All additional information - for example purification method, vectors used, already known sequence peculiarities (e.g. GC- rich regions, duplicated regions, homopolymeric sequences)- are appreciated. They can help to improve the sequencing results since we can adjust the sequencing conditions optimally.

2. Sample concentration/Primer concentration

The amount of template DNA we need for one sequencing reaction (and additional repeated reactions, if needed) is:

BACs/PACs/EBVs 2000ng
Viruses/Phages 1000ng
Cosmids 500ng
Plasmid DNA 300ng
PCR-products (>1kb) 100ng
PCR-products (<1kb) 50ng

Primers supplied by you should have a minimum concentration of 10pmol/µl and optimally an annealing temperature between 50ºC and 60ºC.

3. Shipment of samples

Please send your samples by post in thick-walled tubes and use padded envelopes to avoid damaging of your samples. Do not send your samples as a parcel, this will lead to delays. Refrigeration of the DNA is not necessary. In urgent cases we recommend using a courier service.