Dear community,
In the past, we have sequenced our genomic library pools (Illumina PCR free) in two rounds on the NovaSeq6000. After the first round, we rebalanced the pool based on % PF cluster. We achieved a very good distribution of the individual samples in the pool with a Std dev of 2.
To save a little time, we now tried to achieve the same result with a rebalancing using iSeq. Unfortunately, we achieve very poor results with a Std dev of 9 when we follow the Illumina protocol for rebalancing.
An equimolar pooling of the samples and the inclusion of the index performance via a multiplication factor results in a sample distribution with a Std dev of 5.
How do you rebalance your genome pools and what are your outputs in terms of std dev?
Do you have any idea what we can improve to get back to a lower std dev?
Thanks for your input
Best regards
EliG
In the past, we have sequenced our genomic library pools (Illumina PCR free) in two rounds on the NovaSeq6000. After the first round, we rebalanced the pool based on % PF cluster. We achieved a very good distribution of the individual samples in the pool with a Std dev of 2.
To save a little time, we now tried to achieve the same result with a rebalancing using iSeq. Unfortunately, we achieve very poor results with a Std dev of 9 when we follow the Illumina protocol for rebalancing.
An equimolar pooling of the samples and the inclusion of the index performance via a multiplication factor results in a sample distribution with a Std dev of 5.
How do you rebalance your genome pools and what are your outputs in terms of std dev?
Do you have any idea what we can improve to get back to a lower std dev?
Thanks for your input
Best regards
EliG