Hi Astrid

When you do the blast between the assembled sequence and reference sequence
both sequences need to be in single fasta format (i.e. not multiple fasta).
The assembled sequence is likely to be in a multiple fasta format, so you
will need to load that into Artemis first and save it out as a single fasta
(from the menu: File->Write->All Bases->FASTA format). Then repeat the blast
comparison using this and the coordinates for the blast hits will appear
correctly in ACT along all scaffolds.

I am not sure why your scaffolds are ordered by size and how likely that is
for your assembled sequence.

Regards
Tim

On 1/5/12 9:36 AM, "Astrid von Mentzer" <[email protected]> wrote:

> Hi, 
> 
> I have an assembled sequence (Velvet) which I have compared to a reference
> sequence in ACT. I have run a blastall between the sequences. When I view the
> comparison in ACT, the scaffolds are ordered in size with the biggest first
> and only the biggest scaffold is compared to the reference sequence.
> 
> What I wonder first is: Why are the scaffolds ordered according to size? How
> do I make a comparison between all scaffolds (the whole sequence) to the
> reference strain and not just one scaffold?
> 
> I am new to this so please describe in detail so I understand.
> 
> Cheers, 
> Astrid
> 
> 
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