Whole blood input requirements for Illumina DNA PCR Free

Does the Illumina PCR-Free kit have all the same flexibility using different blood storage tubes as the Illumina DNA Prep (M) Tagmentation workflow? Yes, the Illumina DNA PCR-Free kit is based on the same technology as Illumina DNA Prep, (M) Tagmentation (formerly Nextera DNA Flex) and has similar compatibility.

Which blood tubes have been tested? This prep is optimized for the use of K2-EDTA blood collection tubes from Becton Dickinson. Other blood tubes may work, but they have not been validated.

Is the blood collection kit the same between Illumina DNA Prep, (M) Tagmentation (formerly Nextera DNA Flex) and this kit? If not, can blood collected for Illumina DNA Prep be used with this library prep kit? If so, are there deviations to make to the extraction or library prep protocol? No, these are different products and must not be used interchangeably.

What is the appropriate temperature for storage and transport of blood tubes? Store and transport at 2-8°C. Perform lysis reactions within 3 days of blood being collected for optimal results.

What happens if blood collected >3 days ago is used? Yields and library preparation efficiency can vary.

Can frozen blood be used? Frozen blood is not supported.

What happens if more or less than 20 uL of blood is 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.

For any feedback or questions regarding this article (Illumina Knowledge Article #3433), contact Illumina Technical Support [e-mail].