Question: Can I split my library pool into two 0.2 ml tubes in the SPRIselect Library Concentration Protocol for the Targeted Gene Expression assay?
Answer: When using our Demonstrated Protocol for SPRIselect Library Concentration, it is possible to split the library pool into two tubes if the volume exceeds that which can fit in one 0.2 ml tube.
For example, if the volume of the library pool + Cot DNA is 120 µl, you would need to add 216 µl SPRIselect reagent (1.8X) for a total volume of 336 µl. This would exceed the volume that can fit in one 0.2 ml tube. In this case, the sample can be split into two 0.2 ml tubes, as outlined here:
- Pool libraries and Cot DNA
- Split into two 0.2 ml tubes
- Add 1.8X SPRIselect reagent to each.
- Wash as outlined in the Demonstrated Protocol
- Elute each pellet by adding 9.5 µL of Modified Hybridization Mix to each tube
- Collect 8.5 µL of supernatant from each tube
- Pool supernatants together for a total volume of 17 µL
- Proceed with library hybridization
It may be beneficial to add an extra PCR cycle to the post-capture library amplification step to account for the loss of material due to using lower elution volumes per SPRI pellet.
Note: The Targeted Gene Expression assay has been discontinued. For alternative options, please see: What options are there for performing a targeted enrichment for my gene expression libraries?
Product: Targeted Gene Expression