BLT PF handling for the Illumina DNA PCR Free workflow questions

Why does the BLT-PF tube have to stay upright in storage? Illumina recommends that the BLT-PF stay upright in storage to enable easy resuspension and avoid bead drying. Make sure beads are thoroughly mixed and resuspended prior to use.

If there bead residue that dried inside the BLT-PF tube, is it a problem? No, proceed as normal, but try to resuspend by inversion and vortexing.

How to proceed if too much or too little BLT-PF is added to a sample? Using less or more of BLT-PF will impact library yield and size distribution. If various amounts of BLT-PFs were used to create libraries, Illumina recommends individually quantifying the libraries before pooling. Libraries may still be sequenced, though certain sequencing metrics may be affected.

How to proceed if the samples with BLT-PF are left on the magnet longer than is stated in the protocol? Leaving BLT-PF on the magnet for several extra minutes does not lead to any downstream issues with library prep; however, Illumina does not recommend leaving the library prep for long stretches of time (>10 minutes).

What happens if BLT-PF beads dry? Overdried BLT-PF beads can reduce library yield.

What happens if the beads precipitate out of solution?

Resuspend beads and proceed as normal.

For any feedback or questions regarding this article (Illumina Knowledge Article #3454), contact Illumina Technical Support [e-mail].