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In order to sequence monotemplates (where most sequenceable fragments have exactly the same sequence), it is often necessary to significantly dilute or mix the sample with a complex genomic library to enable registration of clusters.
In order to sequence a coelacanth's (Latimeria chalumnae) genome, scientists required fresh tissue and blood.
An improved genome Walking approach was used in order to sequence the remaining adjacent genes of the cluster.
In order to sequence a huge number of samples in a single sequencing run, a stable and reliable method for identifying the generated reads is required.
Firstly, template read-length is sacrificed in order to sequence the barcode, although the read length can be extended if required.
Each preparation was linked to a bar-coded adapter in order to sequence the two libraries in the same channel, thus avoiding any undesired bias and obtaining two independent datasets that could be further compared.
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All PCR products were cloned prior to sequencing in order to ensure sequence accuracy.
Also, in Illumina sequencing, DNA from the PhiX phage is often added to the sequencing reaction, in order to calibrate sequence quality scores.
In order to characterise sequence differences in detail, multiple sequence alignments (MSAs) were generated on the basis of a representative selection of TrpA and TrpB sequences.
In order to completely sequence each homolog, 5'-RACE and primer walking sequencing were performed.
In order to survey sequence variation in 5S-NTS, random clones, 7 (Paphiopedilum niveum) or 8 (all others) per species, were sequenced (Additional file 1).
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