Real-Time
(DNA) Real-time Exicycler 96 Rotor-Gene Q/6000 IU
Mix1 Mix2 IU/μl IU/μl IU/μl IU/μl IU/μl IPC NTC C 1 Lot# 2 Freeze & thawing
1 MSDS: Material Safety Data Sheets
Real- (TaqMan time ' FAM ' BHQ1 ' BHQ1 ' JOE in vitro in vitro 1 Validation 2 Conserved
Real-Time Real-time Real-time Real-time SYBR Green DNA DNA DNA 1 Post 2 UV Transilluminator 3 Non Specific Methods 4 Signal
TaqMan Real-time MRD 1 Specific Methods 2 Minimal Residual Disease
(IU) IU/ml IU/l IU WHO WHO 1 Herpesviridae 2 IU: International Unit 3 WHO: World Health Organization 4 Viral Loads 5 1 st WHO international Standard for Human Cytomegalovirus for Nucleic Acid Amplification Techniques, code: 09/162
pvalue IU/l IU/l CMV IU/l 1 Analytical Sensitivity
Human Herpes Simplex virus type 1 Human Herpes Simplex virus type 2 Varicella Zoster Virus 2nd WHO International Standard for Hepatitis B DNA for Nucleic Acid Amplification (NAT) Assay Epstein-Barr Virus 2nd WHO International Standard for Parvovirus B19 DNA for Nucleic Acid Amplification (NAT) Assay WHO 2nd International Standard For Hepatitis C Virus RNA for Genomic Amplification Technology Assays Human Papilloma Virus Type16 Human Papilloma Virus Type18 (DNA) 1 Analytical Specificity 2 Cross Reactivity
IU/µl IU/µl IU/µl CMV. IU/µl IU/µl 1 Linear range 2 Precision 3 Variance 4 Intra assay variation 5 Inter assay variation 6 SD: Standard Deviation
. C t IU/µl Ct Ct CV Ct IU IU CV IU Intra Assay variation Inter Assay variation Total variance Exicycler96 Ct Ct CV Ct IU IU CV IU Intra Assay variation Inter Assay variation Total variance Exicycler96 1 CV: Coefficient of Variation 2 C t : Cycle of threshold
Ct Ct CV Ct IU IU CV IU Intra Assay variation Inter Assay variation Total variance Rotor Gene Q Ct Ct CV Ct IU IU CV IU Intra Assay variation Inter Assay variation Total variance Rotor Gene Q
CMV. 1 Diagnostic Evaluation 2 Clinical Sensitivity 3 Clinical Specificity
Real-time (Heating-Block) DNA rpm Real-time 1 DNase RNase Free 2 Vortex 3 Spin 4 Strip 5 Polymerase Chain Reaction ()
EDTA g 1 E.D.T.A
C C C mg/dl mg/dl IU/ml
dntp Mix1 Mix2 IU R 2 R^2 e IPC) C t 1 Efficiency (e ) 2 Internal Positive Control
Real-time Real-time CMV N.T.C FAM CMV JOE C t 1 No Template Control
DNA Roche (Cat#) 11 858 874 001 DNA/RNA DynaBio Viral Nucleic Acid Extraction Mini Kit High Pure Viral Nucleic Acid Kit DNA Elution Buffer 1 Cross Contamination 2 Multiplex
Real-time 1 Sampling 2 Under or Over estimation 3 Duplicate
x x Real-time 1 Run
Exicycler96 CMV (Scan) FAM JOE RotorGene 6000 RotorGene Q CMV (Scan) FAM JOE
ExiCycler96 Exicycler Run Design Experiment File a. Master CMV b. Protocol Takapouzist c. Plate 920101 /.,
Append Incubate Temperature: 95 Temperature: 95 Temperature: 55 Time: 5 min Append Time: 15 sec Append Time: 50 Sec Append Temperature: 72 a. Line=2 b. Cycle=50 Append Scan Incubate Time: 20 Sec Append Go to
JOE BHQ FAM BHQ Append Plate Add Probe OK.1.2 B5.3
E5 B5 E5 Probe Name Assign.4 Type Sample Name Assign Sample Name.5
OK Plate Protocol Save Exicycler3 Run Master save as
.8 Run Exicycler Master Design Experiment (open) Door Run.9.11.11 OK
RotorGene Q RotorGene 6000 New Locking Ring Attached New Quick Start 36-Well Rotor Next Next
Gain Optimisation Auto-Gain Optimisation Set Up OK Green Auto-Gain Optimisation Set Up Add
Add Target Sample Range Ok Yellow Auto-Gain Optimisation Set Up Add
Target Sample Range Ok Green Perform Optimisation Before 1 st Acquisition Close
Next Start Run
RotorGene 6000 IU/µl RotorGene Q
File Exicyler96 Exicycler Run Exicycler Analysis Open Open Absolute Quantification Ok
Trend Average Sub. Method Threshold Level Flu. Graph FAM JOE STD Curve R^2 Efficiency Ct Report Form
RotorGeneQ RotorGene6000 Analysis Analysis Show
Dynamic Tube Slope Correct Auto Scale Linear Scale Threshold Maximize
Quantitation Analysis Ct Standard Curve R^2 Efficiency Quant. Results JOE
(DNA) FAM CMV JOE FAM JOE FAM C t CMV JOE FAM 1 Flowchart 2 Trouble Shooting
JOE JOE ± FAM FAM R^2> Ct FAM CMV JOE IU/ml
IU IU IU/l (WHO) Result (IU/ml) = Result (IU/µl) Elution Volume (µl) Sample Volume (ml) IU/µl (IU/ml) = IU/ml R^2 e 1 PBS
FAM CMV JOE FAM Symptom FAM JOE ± Ct DNA FAM
Symptom FAM Real-time
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