With primer walking we sequence very long stretches of DNA (PCR fragments, plasmids, BACs, Cosmids, Adenoviruses…) by aligning more reactions triggered by different oligos. Sequence reactions are done by drawing oligos every 400 bases , designed either on a reference sequence or on the result of previous reactions.
ssDNA:
You must: Prepare the DNA and specify how many bases you want to sequence. Send us, if you have it, a reference sequence that has a good homology with the one you want to sequence. For the quantities to be sent, agree with the laboratory.
We must: Synthesize oligos every 400bp and create your own personal oligo bank (duration two years) by activating an interactive folder in the reserved area.
Align all the readings made and send you the consensus in fasta format, the single electropherograms and a pairing map of the oligos. The accuracy of the expected result is 99.8%.
Agreements and strategies can vary depending on the complexity of the sequencing that can be encountered.
dsDNA:
You must : Prepare the DNA and specify how many bases you want to sequence. Send us, if you have it, a reference sequence that has a good homology with the one you want to sequence. For the quantities to be sent, agree with the laboratory.
We must : Synthesize the for and rev oligos every 400bp and create your own personal oligo bank (duration two years) by activating an interactive folder in the reserved area.
Align all the readings made and send you the consensus in fasta format, the single electropherograms and a pairing map of the oligos. With readings on both strands the accuracy of the predicted result increases up to99.99%.
Agreements and strategies can vary depending on the complexity of the sequencing that can be encountered.
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