Journal: Radiation research

Article Title: NATO BIODOSIMETRY STUDY

doi: 10.1667/RR3236.1

Figure Lengend Snippet: Methods Used by Contributing Laboratories

Article Snippet: PCR with β-actin primer and HotStar MasterMix (Qiagen), 30 cycles High capacity cDNA archive kit, AB 1×/25°C/10 min, 1×/37°C/ 120min 18SrRNA-CT TaqMan Universal Master Mix, AB GADD45A, Hs00169255_m1 BAX, Hs00180269_m1 DDB2, Hs00172068_m1 CDKN1A, Hs00355782_m1 1×/50°C/2 min 1×/ 95 °C/10 min 40×/ 95 °C/1 min 60°C/1 min GeneAmp 7900 AB automated 18S rRNA ΔΔ CT approach yes// yes// yes// yes 5 QIAamp RNA Blood Mini Kit RNase-free DNase-Set (Qiagen) RNAse-free water No// yes//yes PCR Real-Time with adapted primers (single copy costitutive gene) High capacity cDNA archive kit, AB 1×/25°C/10 min, 1×/37°C/ 120min 18SrRNA-CT TaqMan Universal Master Mix, AB GADD45A, BAX, DDB2, CDKN1A 1×/50°C/2 min 1×/ 95 °C/10 min 40×/ 95 °C/1 min 60°C/1 min StepOnePlus ABI automated 18S rRNA ΔΔ XT approach yes// yes// yes// yes 6 QIAamp RNA Blood Mini Kit RNase-free DNase-Set (Qiagen) TE or RNAse- Free water No, agarose gel//yes Nano- Drop//yes// Yes//-RT control High capacity cDNA archive kit, AB 1×/25°C/10 min, 1×/37°C/120 min 1×/85°C/5 min HPRT1-CT TaqMan,PerfeCTa ® MultiPlex qPCR SuperMix, Quanta bioscience PCNA, PHPT1, TIGAR, CCNG1, DDB2, FDXR, GADD45A, MDM2 1×/95°C/2 min 40× /95°C/10 s 60°C/1 min Rotor-Gene 6000 Qiagen fixed HPRT1 2 Standard curves relative quantification yes//yes// yes// HPRT1, (-)RT and NTC 7 no RNA Isolation- direct from stabilized blood testing with NEAT-CLPA assay no no no no cDNA synthesis NA NA Finnzymes QPCR F-450 30-plex assay, which includes markers for PCNA, DDB2, BAX, FDXR, GADD45A and several other proprietary markers.

Techniques: Isolation, Concentration Assay, SYBR Green Assay, RNA Extraction, Software, Multiplex Assay, Electrophoresis

Journal: Radiation research

Article Title: NATO BIODOSIMETRY STUDY

doi: 10.1667/RR3236.1

Figure Lengend Snippet: Methods Used by Contributing Laboratories

Article Snippet: 3 min, 95°C (RT inactivation priming with optimized mix of oligo(dT) and random oligomers 5′ –3′ assay Rotor-Gene SYBR Green PCR Kit (QIAGEN) Rotor-Gene Probe PCR Kit (QIAGEN) SYBR Green Assays (Qiagen): Hs_GADD45A_1_SG Hs_BAX_3_SG Hs_DDB2_1_SG Hs_CDKN1A_1_SG TaqMan Probe Assays (Qiagen): Hs_GADD45A_1_QF Hs_BAX_2_QF Hs_DDB2_1_QF Hs_CDKN1A_1_QF Hs_RN18S1_2_QF SYBR Green: 1 × 5 min 95°C 40 × 5 s 95°C/10 s 60°C TaqMan Probes: 1 × 3 min 95°C 40 × 3 s 95°C/10 s 60°C Rotor-Gene Q (QIAGEN) variable 18S rRNA REST software with efficiency correction yes// yes// yes// no 4 QIAamp RNA Blood Mini Kit RNase-free DNase-Set (Qiagen) RNAse-free water yes//yes, Nano-Drop//yes// yes// convent.

Techniques: Isolation, Concentration Assay, SYBR Green Assay, RNA Extraction, Software, Multiplex Assay, Electrophoresis