Nextera purification

[fraca]

Hello everybody,
I have one question about the nextera kit. If I want to use the kit to do only one or two libraries, so I don't work in plate. I can use other consumables and other kits for the purification (Agencourt AMPure XP 60 ml kit, ZymoTM Purification Kit Catalog)? Have someone try with other kits?
Thanks

[JPC]

Hi, the Nextera_XT kit already has Ampure beads for the clean up, I've used this on the MiSeq with no problems.

JPC

[kcchan]

When did Illumina start including the Ampure XP beads? I thought they only include the library normalization beads in their Nextera XT kit.

[RCJK]

For small sample number with Nextera kits we use the Zymo columns instead of the plates. We still use Ampure XP beads for cleanups.
The beads in the Nextera XT kit are for normalization, not Ampure XP beads.

[JPC]

When did Illumina start including the Ampure XP beads? I thought they only include the library normalization beads in their Nextera XT kit.

I'm not sure when they were added as this is the first protocol I have used on the MiSeq

-Nextera XT Sample Preparation Guide 15031942. Pages 30/31

[kcchan]

I'm not sure when they were added as this is the first protocol I have used on the MiSeq

-Nextera XT Sample Preparation Guide 15031942. Pages 30/31

According to that page (Part # 15031942 Rev. C Page 30), the AMPure XP beads are still supplied by User. They are not included with the kit. I believe that only the TruSeq PCR free kits include the DNA purification beads in the box.

[fraca]

Thanks for the answers,
I will use the protocol kits, Instead of the plates version I will use the preps/columns because I have few samples to process. The buffers are the same.

[Joltex]

For small sample number with Nextera kits we use the Zymo columns instead of the plates. We still use Ampure XP beads for cleanups.
The beads in the Nextera XT kit are for normalization, not Ampure XP beads.

When you use the Zymo columns instead of the plates, do you use the volumes and spin speeds specified in the Zymo column protocol (https://www.zymoresearch.com/downloa...35/d4003i.pdf)? Or do you stick with the Nextera protocol?

[fraca]

Hi,
I use the volume and spin of the of the miniprep tubes, and eventually I resuspend in RBS buffer.