New kit: Nextera DNA-Flex

[Genetic Librarian]

Hi,
I am sure many of you got the newsletter from Illumina today.
So, apparently the New DNA Flex kit is an all-in-one solution for Nextera XT, Nextera and TruSeq DNA Nano Users.
Main benefit seems to be its tolerance towards input concentrations (so no more super accurate Qubit needed). Main disappointment is the separately sold index set with yet again no full unique dual indexing capability (but at least a columnwise start). They missed a chance here for a really fresh start.

I would be happy to read about your first test results here. We will at least give it a try.

[nucacidhunter]

I wonder if you know how its price compares to standard Nextera or Nextera XT.

[Genetic Librarian]

Prices are already up on their website.
Per sample it is more expensive than Nextera XT and TruSeq Nano, but cheaper than Nextera (at least in EURO). So if you use all three kits with bias to Nextera, you would go cheaper, if you mainly use XT, you would pay more.
It needs to be tested how much money you save by not needing to Qubit and savings in hands-on-time.

[nucacidhunter]

Thanks. Input DNA still needs to be quantified to decide on PCR cycles, though accurate normalization is not necessary.

[nucacidhunter]

Facts gleaned from Illumina data sheets:
1- Input DNA quantification is required but not accurate normalisation
2- DNA quality will affect the library as does with other Nextera kits and not suitable for FFPE or cfDNA
3- Less flexible for modifying library insert size
4- Comparisons are to Nextera kits or Nano not leading kits that outperform Nano kit
5- Data sheets does not have extensive quality metrics (lacks uniformity of GC coverage information, duplicates for certain read number)
6- Uniform insert size refers to sequenced library fragments not the actual library insert size

Generally, it is less likely to be considered as a technological advance as other supplier’s kits with one tube fragmentation, combined end repair and A tailing followed by ligation are faster or require similar hands on time with the advantage of lower cost and less bias.

[huguesparri]

The kit includes so-called "SPB" (Sample purification Beads) so you don't need AMpure XP anymore (except for an initial hypothetic sample purification).
It might not justify completely the higher price (when compared to Truseq nano and Nextera XT) but it is still to be taken into account...

[lmj]

We have tested this kit sequencing bacterial whole genomes with the aim of trying to get improved read distribution compared Nextera XT.

Thoughts on the kit;
1) Super easy to use
2) No normalisation required and distribution of reads across different samples in the library much more even
3) Distribution of reads across the genome is better than Nextera XT, still not perfect but much better
4) While slightly more expensive that NexteraXT I think it actually comes out a but cheaper as you don't need AmpureXP beads and you don't need to run gels on individual samples before pooling.

For us the flex kit is an improvement over Nextera XT

[lac302]

The bead based normalization used in the XT kits have always worked well in our lab and we've always normalized inputs (1ng) using a nanodrop. Considering the Flex kit but have a few questions for those that have used it.

Is the improved read distribution stated in the Flex marketing material due to less tagmentation bias in comparison to the XT kit?

Also does anyone have any experience with how even or uneven the library concentration is for each sample when using <100ng starting material?