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Old 09-21-2018, 09:43 AM   #1
UCan'tBcereus
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Location: New England

Join Date: Aug 2018
Posts: 5
Default Nextera Flex 1/2 and 1/4 Reaction Experiences?

Hi Everyone,

We are about to run through a test of the Nextera Flex kit, and I wanted to see if anyone has had a positive experience cutting down the RXN volume down to 1/2 RXN or 1/4 RXN.

When we use Nextera XT, we cut down to 1/2 RXN and 1/4 RXN successfully with no issues, other than having to experiment a little with DNA input.

I have full confidence that Nextera Flex libraries will work at 1/2 RXN and 1/4 RXN, but I was wondering if anyone has seen how the "no normalization required with >100 ng DNA input" aspect holds up. I.e at 1/2 RXN, has anyone seen that at 50 ng DNA input the library yields are close enough to not require extensive normalization?

Any other Nextera Flex tips that anyone has noticed would be great too. Thanks everyone!

Quick Side Note: After talking to Illumina one of their reps mentioned that 96 Indexes (UDI not combinatorial) will be released sometime next month for Nextera Flex, with 384 UDI indexes being available "early 2019", in case anyone was interested.
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Old 09-23-2018, 04:34 PM   #2
ikripp
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Location: QLD, Aus

Join Date: Jan 2018
Posts: 16
Default

We do 1/5 reactions for Flex as we also did for XT before changing over.
They appear to be comparable and we have had no issues.
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Old 10-02-2018, 11:11 AM   #3
thang198
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Location: USA

Join Date: May 2018
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Default

Quote:
Originally Posted by UCan'tBcereus View Post
Hi Everyone,

We are about to run through a test of the Nextera Flex kit, and I wanted to see if anyone has had a positive experience cutting down the RXN volume down to 1/2 RXN or 1/4 RXN.

When we use Nextera XT, we cut down to 1/2 RXN and 1/4 RXN successfully with no issues, other than having to experiment a little with DNA input.

I have full confidence that Nextera Flex libraries will work at 1/2 RXN and 1/4 RXN, but I was wondering if anyone has seen how the "no normalization required with >100 ng DNA input" aspect holds up. I.e at 1/2 RXN, has anyone seen that at 50 ng DNA input the library yields are close enough to not require extensive normalization?

Any other Nextera Flex tips that anyone has noticed would be great too. Thanks everyone!

Quick Side Note: After talking to Illumina one of their reps mentioned that 96 Indexes (UDI not combinatorial) will be released sometime next month for Nextera Flex, with 384 UDI indexes being available "early 2019", in case anyone was interested.
Hi UCan'tBcereus,
Would you please tell me how much input you used for the 1/2 and 1/4 RXN?
Thank you,
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