PHASIFY MAX cfDNA Extraction Kit
The PHASIFY MAX cfDNA Extraction Kit (CE, NMPA-certified) isolates and concentrates cfDNA from plasma, demonstrating significantly improved recovery against standard solid phase extraction.
- For in-vitro diagnostics use only.
- High Yield: Multiple fold increase in cell-free DNA recovery compared to solid phase extraction
- High Concentration: Flexible elution volume as low as 5 μL
- Simplicity of Operation: No specialized equipment such as magnetic racks and vacuum pumps
|Applications||ddPCR, PCR, real-time PCR, NGS, other|
|Time per run||~180 mins|
|Input volume||1 mL, 2 mL|
|Elution volume||5 µL - 150µL|
|Shelf life stability||12 months|
|Intended use||In-vitro diagnostics use only.|
|Shipping condition||Room temperature|
RT Box: Stored at 15-30°C
COLD Box: Stored at 4°C
Figure 1- Improved yield of circulating tumor DNA (ctDNA)
The cfDNA was extracted from 89 tissue positive cancer patient plasma samples (1 mL) using PHASIFY MAX and the Qiagen QIAamp kit. Total DNA recovery was measured using Quant-iT™ PicoGreen™ dsDNA Assay Kit. On average PHASIFY™ MAX demonstrated a 3.8-fold increase in total DNA recovery compared to Qiagen QIAamp. (P-Value <0.0001 Wilcoxon Signed-Rank). Note, 2 outliers were excluded in which PHASIFY™ MAX generated 209-fold and 800-fold greater recovery.
Figure 2 – Improved mutation detection
In 13 samples, mutations were detected in samples extracted with PHASIFY™ MAX, but not with Qiagen QIAamp.
Figure 3 – Greater Recovery of Desired cfDNA Fragments
DNA was extracted from 1 mL healthy human plasma using PHASIFY™ MAX or the Qiagen QIAamp Circulating Nucleic Acid kit. The extracted DNA was analyzed on Agilent Bioanalyzer 2100, where 35bp
and 10,380bp peaks correspond to internal markers.
If you have less plasma input than the volume indicated on the kit, we recommend bringing the sample volume up to the indicated volume by adding 1x PBS (pH 7.4) prior to step 1. However, we do not recommend using less than 80% v/v of plasma.
A higher volume input might disrupt phase forming. We recommend using the plasma input volume as indicated on the kit.
Under controlled conditions, the bottom phase will be approximately 800-860 μL for the 1 mL plasma input kit and 900-950 μL for the 2 mL plasma input kit. Regardless of the volume, we recommend extracting all the bottom phase during Step 5.
This will depend on your downstream application of the extracted DNA. It is possible that any transfer of the wrong phase can impact your results. We have found that transferring small amounts (<10% by volume) will not significantly impact results for most applications, however getting extra bottom phase in step 7 would affect performance of size selection. We recommend to avoid transfer of the wrong phase in all cases.
We recommend transferring only 120 μL of the red top phase. Transferring more of the top phase may result in higher recovery/yield, but it increases the risk of accidentally pipetting and transferring contaminants from the bottom phase. We recommend extracting the specified volume of the top phase.