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Old 10-30-2013, 03:05 AM   #1
Junior Member
Location: Bel

Join Date: Jan 2013
Posts: 4
Default Nimblegen whole exome capture, hybridisation step

We are currently performing lib prep using nimblegen's SeqCap EZ whole exome capturing kit.

At the hybridisation step in the protocol, we have to incubate the samples at 47C for 64-72 hours.

For practical reasons, we can only continue the protocol after 4 days instead of the proposed 3.

We have several options:

1) hybridise the sample for 96 hours
2) hybridise samples for 72 hours and keep them at 4C untill we are ready to proceed
3) postpone the hybridisation step

As we want to sequence the samples ASAP, we prefer option 1 or 2.

Does anyone have experience with this? Please share.
kodbeeck is offline   Reply With Quote
Old 11-13-2014, 11:50 PM   #2
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Location: Brussels

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Good question. I have the same issue. Which option did you eventually do? I'm thinking to do a hybridization of 92 hours. This shouldn't give any problems, right?
JeroenD is offline   Reply With Quote
Old 02-15-2015, 06:36 AM   #3
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Location: Saint Louis

Join Date: Feb 2015
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Any problems with doing hybridization longer? 90 hours?
herter is offline   Reply With Quote
Old 04-28-2015, 03:00 AM   #4
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Location: Barcelona

Join Date: Nov 2011
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Default any expierience

...same here. Looking eagerly for an answer.
Thank you
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Old 04-28-2015, 04:13 AM   #5
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Location: Saint Louis

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Nimblegen rep confirmed that it can go longer, just make sure that nothing can evaporate.
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