WHAT CAN YOU DO TO OBTAIN
OPTIMAL SEQUENCING RESULTS?
1. DNA preparation
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
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.