Using dried blood spots as input into the Illumina DNA PCR Free library prep kit questions

Can blood spots on cards (dried blood spots, DBS) be used? Yes, see the Reference Guide for details.

Which blood spot cards have been tested? Prep is optimized for the use of Whatman 903. Other blood spot cards may work, but they have not been tested.

What is the appropriate temperature for storage and transport of dried blood spots? Dried blood spots are to be shipped and stored at room temperature, in a sealed bag containing desiccant packs.

What happens if more or fewer than 6 punches are added to the lysis reaction? This is not recommended as it may affect the efficiency of the lysis reaction, which will ultimately impact the amount of DNA obtained and, therefore, the final library yield. Discoloration will also become evident in the eluted DNA. Using fewer than 6 punches will generate libraries in most cases, but the low input protocol is recommended, and DNA will not saturate the BLT-PF (libraries will not be normalized).

Can the Illumina Purification Beads (IPB) purification at the end of the integrated extraction be removed? No, the IPB purification required to remove contaminants prior to library preparation.

Can the volume of IPBs be increased to recover more DNA?

Illumina recommends following the protocol as written, otherwise there will be loss of yield and potential contaminant carryover.

For any feedback or questions regarding this article (Illumina Knowledge Article #3436), contact Illumina Technical Support [e-mail].