03/11/22
The Illumina DNA PCR-Free Library Prep, Tagmentation kit offers multiple workflows to accommodate variations in sample type, DNA input amount, and application. See the following table for considerations when choosing the optimal protocol. Always consult the Reference Guide for full details, including validated catalog numbers.
Equipment/consumables considerations
| Input Amount/Type | 25–99 ng DNA | 100–299 ng DNA | 300–2000 ng DNA | Whole blood/Dried blood spots/Saliva | |
|---|---|---|---|---|---|
| Protocol compatibility | Thermal cycler, Low input | Thermal cycler, Standard input (sub-saturation) | Thermal cycler, Standard input(saturation) | Hybex (saturation) | Thermal cycler, Standard input(saturation) |
| Incubation steps method | Thermal cycler | Thermal cycler | Thermal cycler | Hybex + MIDI heat block insert | Thermal cycler |
| Type of magnet required | DynaMag-96 Side Skirted Magnet; Thermo Fisher Scientific, #12027 | DynaMag-96 Side Skirted Magnet; Thermo Fisher Scientific, #12027 | DynaMag-96 Side Skirted Magnet; Thermo Fisher Scientific,#12027 | Magnetic Stand-96 Thermo Fisher Scientific, #AM10027 | DynaMag -2 Magnet (Thermo Fisher Scientific, #12321D) and DynaMag-96 Side Skirted Magnet (#12027) |
| Plate type | PCR | PCR | PCR | MIDI | PCR |
| Mixing method | Pipetting | Pipetting | Pipetting | Shaking | Pipetting |
| Additional equipment required | Real-time PCR detection system | Real-time PCR detection system | Qubit or Real-time PCR detection system | Shaker (Bioshake XP or Bioshake IQ); Qubit or Real-time PCR detection system | Additional equipment depends on type on input material used; see reference guide |
| Additional Illumina reagents* | Not applicable | Not applicable | Not applicable | Not applicable | Illumina Lysis Reagent Kit |
*Note: most instruments and sequencing consumables require additional custom sequencing primers. See the Custom Primer Requirements for the Illumina DNA PCR-Free Prep, Tagmentation Kit bulletin for details.
Quantification considerations
| Input Amount/Type | 25–99 ng DNA | 100–299 ng DNA | 300–2000 ng DNA | Whole blood/Dried blood spots/Saliva |
|---|---|---|---|---|
|
Quantification of Input DNA Required |
Yes | Yes | No | No |
|
Quantification of final individual libraries required |
Yes | Yes | No | No |
|
Quantification of final library pool required |
No | No | Yes | Yes |
|
Quantification method for individual libraries |
Kapa qPCR, replicates required | Kapa qPCR | None | None |
|
Quantification method for final library pool |
None | None | ss Qubit* or Kapa qPCR | ss Qubit* or Kapa qPCR |
* ss Qubit = single-stranded Qubit reagents
|
Input Amount/Type |
25–99 ng DNA | 100–299 ng DNA | 300–2000 ng DNA | Whole blood/Dried blood spots/Saliva |
|---|---|---|---|---|
|
NovaSeq 6000 loading method |
XP |
XP could be required, depending on final yield of libraries |
XP or standard | XP or standard |
| Size selection | Single-sided | Double-sided | Double-sided | Double-sided |