Reducing costs in library preparation

F.Lazaro · 07-26-2017, 03:25 AM

Hello all,

I have read in other posts that is feasible to reduce volumes for library preparation when using the Nextera XT kit. Do you know if its also posible to reduce the volumes by a half when using other kits (e.g. Nebnext Ultra, TruSeq Nano...)

nucacidhunter · 07-27-2017, 02:44 AM

With TreSeq and Nebnext and maybe others as well it is Ok. The downside could be reduced library diversity but that is only important if a library is going to be sequenced deeply (more than 200M reads).

Arthur45 · 08-10-2017, 02:23 AM

Hi all,

I wonder if anyone had experience with miniaturisation of NEB Ultra II Prep kit? Has anyone successfully sequenced whole genome with half the volumes of reagents?

Cheers,

arthur

Simone78 · 08-18-2017, 09:04 AM

Hi,
that´s something we are thinking as well. I don´t see any reasons why it shouldn´t work. I mean, the 50 ul reactions recommended in these Illumina kits are ridiculously expensive! That´s why they want you to do it, so you will buy more kits.
As said above, the Nextera reaction has been shown to work as good when performed in few hundreds nanoliter compared to the standard 20 ul recommended by Illumina. This applies to any reaction I have done so far (RT, PCR, tagmentation, etc).
In general, just make sure to decrease the input accordingly if you cut the volumes. I hope this helps!

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07-26-2017, 03:25 AM	#1
F.Lazaro Junior Member Location: Spain Join Date: Jun 2017 Posts: 7	Reducing costs in library preparation Hello all, I have read in other posts that is feasible to reduce volumes for library preparation when using the Nextera XT kit. Do you know if its also posible to reduce the volumes by a half when using other kits (e.g. Nebnext Ultra, TruSeq Nano...)

07-27-2017, 02:44 AM	#2
nucacidhunter Jafar Jabbari Location: Melbourne Join Date: Jan 2013 Posts: 1,238	With TreSeq and Nebnext and maybe others as well it is Ok. The downside could be reduced library diversity but that is only important if a library is going to be sequenced deeply (more than 200M reads).

08-10-2017, 02:23 AM	#3
Arthur45 Junior Member Location: London Join Date: May 2017 Posts: 7	Hi all, I wonder if anyone had experience with miniaturisation of NEB Ultra II Prep kit? Has anyone successfully sequenced whole genome with half the volumes of reagents? Cheers, arthur

08-18-2017, 09:04 AM	#4
Simone78 Senior Member Location: Basel (Switzerland) Join Date: Oct 2010 Posts: 208	Hi, that´s something we are thinking as well. I don´t see any reasons why it shouldn´t work. I mean, the 50 ul reactions recommended in these Illumina kits are ridiculously expensive! That´s why they want you to do it, so you will buy more kits. As said above, the Nextera reaction has been shown to work as good when performed in few hundreds nanoliter compared to the standard 20 ul recommended by Illumina. This applies to any reaction I have done so far (RT, PCR, tagmentation, etc). In general, just make sure to decrease the input accordingly if you cut the volumes. I hope this helps!