Journal: Cancer Medicine

Article Title: Digital PCR improves the quantitation of DMR and the selection of CML candidates to TKIs discontinuation

doi: 10.1002/cam4.2087

Figure Lengend Snippet: Clinical and hematological characteristics of 142 Ph + CML patients with stable DMR comparatively monitored by RT‐qPCR and dPCR grouped in MR class by RT‐qPCR (MR 4.0 vs MR 4.5‐5.0 ) and by dPCR (≥ or <0.468 copies/µL) at enrollment

Article Snippet: We prepared 16 μL of reaction mix containing 8 μL of 2X QuantStudio 3D Digital PCR Master Mix (Thermofisher Scientific), 0.8 μL of 20X TaqMan‐MGB‐FAM‐probe assay, 1.1 μL of diluted cDNA, and 6.1 μL of nuclease‐free water (Qiagen).

Techniques: Digital PCR, Biomarker Discovery

Journal: Cancer Medicine

Article Title: Digital PCR improves the quantitation of DMR and the selection of CML candidates to TKIs discontinuation

doi: 10.1002/cam4.2087

Figure Lengend Snippet: Clinical and hematological characteristics of 111 patients who discontinued TKI treatment comparatively monitored by RT‐qPCR and dPCR grouped in MR class by RT‐qPCR (MR 4.0 vs MR 4.5‐5.0 ) and by dPCR (≥ or <0.468 copies/µL) at enrollment

Article Snippet: We prepared 16 μL of reaction mix containing 8 μL of 2X QuantStudio 3D Digital PCR Master Mix (Thermofisher Scientific), 0.8 μL of 20X TaqMan‐MGB‐FAM‐probe assay, 1.1 μL of diluted cDNA, and 6.1 μL of nuclease‐free water (Qiagen).

Techniques: Digital PCR, Biomarker Discovery

Journal: Cancer Medicine

Article Title: Digital PCR improves the quantitation of DMR and the selection of CML candidates to TKIs discontinuation

doi: 10.1002/cam4.2087

Figure Lengend Snippet: Levels of BCR‐ABL1 transcript measured by dPCR (y‐axis) according to different MR classes calculated by RT‐qPCR (x‐axis). RT‐qPCR BCR‐ABL1 results were grouped into MR 4.0 , MR 4.5 , and MR 5.0 class (“X axis”). When the same samples were analyzed by dPCR, different levels of BCR‐ABL1 copies were measured (“Y axis”). The difference was statistically significant between the MR 4.0 class and the MR 4.5 ( P = 0.01) or the MR 5.0 class ( P = 0.003), but not between the MR 4.5 and the MR 5.0 classes ( P = 0.2). The statistic test used was t test. In box and whiskers plot, center line represents the median values; the boxes’ limits represent the lower and the higher quartile; whiskers define the extreme values; and points represent the single analysis values

Article Snippet: We prepared 16 μL of reaction mix containing 8 μL of 2X QuantStudio 3D Digital PCR Master Mix (Thermofisher Scientific), 0.8 μL of 20X TaqMan‐MGB‐FAM‐probe assay, 1.1 μL of diluted cDNA, and 6.1 μL of nuclease‐free water (Qiagen).

Techniques: Quantitative RT-PCR

Journal: Cancer Medicine

Article Title: Digital PCR improves the quantitation of DMR and the selection of CML candidates to TKIs discontinuation

doi: 10.1002/cam4.2087

Figure Lengend Snippet: Molecular residual disease (MRD) over time as measured by RT‐qPCR and dPCR. (A) MRD monitoring by RT‐qPCR in patients with MR 4.0 at Time Point 0. (B) MRD monitoring by RT‐qPCR in patients with MR 4.5‐5.0 at Time Point 0. (C) MRD monitoring by dPCR in patients with value of BCR‐ABL1 copies/µL ≥0.468 at Time Point 0. (D) MRD monitoring by dPCR in patients with value of BCR‐ABL1 copies/µL <0.468 at Time Point 0

Article Snippet: We prepared 16 μL of reaction mix containing 8 μL of 2X QuantStudio 3D Digital PCR Master Mix (Thermofisher Scientific), 0.8 μL of 20X TaqMan‐MGB‐FAM‐probe assay, 1.1 μL of diluted cDNA, and 6.1 μL of nuclease‐free water (Qiagen).

Techniques: Quantitative RT-PCR

Journal: Cancer Medicine

Article Title: Digital PCR improves the quantitation of DMR and the selection of CML candidates to TKIs discontinuation

doi: 10.1002/cam4.2087

Figure Lengend Snippet: Treatment‐free remission (TFR) curves according to dPCR values. The red curve represents patients discontinued with a dPCR value lower than 0.468 and the black curve patients with a dPCR value higher than 0.468. The probability of maintaining TFR for patients discontinued with dPCR <0.468 was 85% and 83% at 1 and 2 years, respectively. The probability of maintaining TFR for patients discontinued with dPCR ≥ 0.468 was 59% and 52% at 1 and 2 years, respectively. A Kaplan‐Meier analysis was used for the evaluation of TFR. Comparison of subgroups was carried out by a log‐rank test

Article Snippet: We prepared 16 μL of reaction mix containing 8 μL of 2X QuantStudio 3D Digital PCR Master Mix (Thermofisher Scientific), 0.8 μL of 20X TaqMan‐MGB‐FAM‐probe assay, 1.1 μL of diluted cDNA, and 6.1 μL of nuclease‐free water (Qiagen).

Techniques: Comparison

Journal: Cancer Medicine

Article Title: Digital PCR improves the quantitation of DMR and the selection of CML candidates to TKIs discontinuation

doi: 10.1002/cam4.2087

Figure Lengend Snippet: Univariate and multivariate analyses for the prediction of treatment‐free remission (TFR). (A) Univariate Cox regression analysis. Variables included were age at diagnosis, sex, previous therapy with IFN, time to MMR and DMR, Sokal class, type of transcript, time to discontinuation, use of frontline second‐generation TKIs, MR classes considering detectable and undetectable transcript, and dPCR. DMR duration > 5 years (HR 0.2855, CI 95% 0.0931‐0.8760, P = 0.0284) and dPCR (HR 0.2936, CI 95% 0.1302‐0.6618, P = 0.0031) resulted significantly predictable of TFR maintenance. (B) Multivariate analysis—only dPCR retained its significant value (HR 0.2124, CI 95% 0.0637‐0.7082, P = 0.0117)

Article Snippet: We prepared 16 μL of reaction mix containing 8 μL of 2X QuantStudio 3D Digital PCR Master Mix (Thermofisher Scientific), 0.8 μL of 20X TaqMan‐MGB‐FAM‐probe assay, 1.1 μL of diluted cDNA, and 6.1 μL of nuclease‐free water (Qiagen).

Techniques: Biomarker Discovery

Journal: Cell Reports Medicine

Article Title: Pharmacological Activation of Non-canonical NF-κB Signaling Activates Latent HIV-1 Reservoirs In Vivo

doi: 10.1016/j.xcrm.2020.100037

Figure Lengend Snippet:

Article Snippet: RT-dPCR reaction was performed in 20 μL containing 10 μL 2x QuantStudio 3D Mastermix (ThermoFisher), 2 μL Superscript VILO (ThermoFisher), 1 μL Taqman 20x Mastermix containing 900 nM primers and 250 nM probe (Thermofisher), and 200 ng of template RNA with the following cycling conditions: 30 min at 50°C, 10 min at 96°C, 40 cycles each consisting of a 30 sec at 96°C followed by 60°C for 2 min, a final 2 min extension at 60°C, and final hold at 10°C.

Techniques: Virus, Modification, Recombinant, Viability Assay, Multiplex Assay, Digital PCR, Software, Cell Isolation

Journal: Pharmaceutics

Article Title: Impact of Processing Parameters on the Quality of Pharmaceutical Solid Dosage Forms Produced by Fused Deposition Modeling (FDM)

doi: 10.3390/pharmaceutics11120633

Figure Lengend Snippet: Summary of the interactions between the materials properties and the machine and process parameters in a fused deposition modeling (FDM) three-dimensional (3D) printing process.

Article Snippet: The printing head of a MakerBot ® Replicator 2X desktop FDM 3D printer (MakerBot Industries LLC.

Techniques:

Journal: Oncology Letters

Article Title: Clinical significance of BRAF V600E mutation in circulating tumor DNA in Chinese patients with melanoma

doi: 10.3892/ol.2017.7529

Figure Lengend Snippet: A negative sample from a melanoma patient of ctDNA in 3D dPCR (A) and the other positive for this mutation (B). A negative sample in tumor tissue by Sanger Sequencing (C) and the other positive for this mutation (D). In plot of 3D dPCR, yellow cluster corresponds to negative droplets, red cluster corresponds to droplets positive for wild-type, blue cluster corresponds to droplets positive for mutant BRAFV600E, and green cluster corresponds to droplets both positive for wild-type and mutant BRAFV600E. In plot of Sanger Sequencing, the nucleotide refers to 1799T >A and the amino acid refers to V600E. dPCR, digital PCR; BRAFV600E, B rapidly accelerated fibrosarcoma.

Article Snippet: Each 15 µl dPCR reaction contained 0.375 µl BRAF assay, 7.5 µl QuantStudio™ 3D Digital PCR Master Mix 2X (Life; Thermo Fisher Scientific, Inc.), 6.125 µl nuclease-free water (Life, Thermo Fisher Scientific, Inc.), and 1 µl cfDNA.

Techniques: Mutagenesis, Sequencing, Digital PCR