Nimblegen whole exome capture, hybridisation step

[kodbeeck]

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 47°C 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 4°C 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.

[JeroenD]

Hi,

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?

[herter]

Any problems with doing hybridization longer? 90 hours?

[drops]

any expierience

...same here. Looking eagerly for an answer.
Thank you

[herter]

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