Hi everybody,
We plan to do a complete run with 454 sequencing (100mb). I already did a 16th of a run (GS FLX 250pb average) and it has generated only 6000 sequences (I was expected 12000) the compagny I worked with to do the 454 said that it is normal and if we want to do a complete run we will have only 300000 sequences instead of the 400000. I did amplicon sequencing with 9 samples pooled together (with 9 different 4bp tags between the adaptator and the primer). Now I have been in touch with others compagny and they said that I should have 400000 sequences and even more (one compagny reckon that they can achieve 30% more). This compagny suggest that the low nb of sequences from the previous 16th of run may be due to a bad design of the tags (redundancy) and I have effectively some that are redundant but it is not the sample that have the less sequences.
Did anyone run a complete run and can we really have the 400000 sequences promised by roche with GS FLX (250pb read average). Does the design of the tag can really be a problem (I thought 454 was precise at 99.9%).
Thank you for answering.
We plan to do a complete run with 454 sequencing (100mb). I already did a 16th of a run (GS FLX 250pb average) and it has generated only 6000 sequences (I was expected 12000) the compagny I worked with to do the 454 said that it is normal and if we want to do a complete run we will have only 300000 sequences instead of the 400000. I did amplicon sequencing with 9 samples pooled together (with 9 different 4bp tags between the adaptator and the primer). Now I have been in touch with others compagny and they said that I should have 400000 sequences and even more (one compagny reckon that they can achieve 30% more). This compagny suggest that the low nb of sequences from the previous 16th of run may be due to a bad design of the tags (redundancy) and I have effectively some that are redundant but it is not the sample that have the less sequences.
Did anyone run a complete run and can we really have the 400000 sequences promised by roche with GS FLX (250pb read average). Does the design of the tag can really be a problem (I thought 454 was precise at 99.9%).
Thank you for answering.
Comment