Rotor-Gene Q PCR Sample to Insight
PCR Rotor-Gene Q PCR DN QIGEN PCR Rotor-Gene Q QIGEN Rotor-Gene Q Rotor-Gene Q PCR High Resolution Melt SNP QIGEN Rotor-Gene Q QIGEN PCR HRM High Resolution Melt DN 8 12 mirn 2 Rotor-Gene Q 7/215
Rotor-Gene Q PCR 1 2 ±.2 ±.2 1/2 ROX.6.5.4.3.2.1. 5 15 25 35 4 2 1.6.5.4.3.2.1 3. 5 6 1 Reaction chamber 7 2 Tubes in rotor spin past optics 3 Motor spindle assembly 4 4 LED lights 5 Lens 6 Emission filters 7 Photomultiplier detector 5 15 25 35 4.8.6.4.2. 1 2 3 4 Centrifugal fan drives cool air into chamber Vent opens to expel hot air Centrifugal fan drives air around chamber Heating element switches off 2. PCR DN 2 3 ng.6 ng 1, 2 PCR IL1R2 TaqMan Rotor-Gene Probe PCR Kit Rotor-Gene Q 5 replicate C T 1.7 CL2 TaqMan Rotor-Gene Probe PCR Kit Rotor-Gene Q ROX DN PCR 72 replicate C T 24.94.5 CV.2 1. Rotor-Gene Q Rotor-Gene Q 7/215 3
Rotor-Gene Q SYR Green TaqMan FRET UV 6 1 Rotor-Gene Q 1. Rotor-Gene Q 72-Well Rotor nm nm lue 365 ± 2 46 ± 2 Marina lue, Edans, othell lue, lexa Fluor 35 Green 47 ± 1 51 ± 5 FM, SYR Green I, Fluorescein, EvaGreen, lexa Fluor 488 Yellow 53 ± 5 557 ± 5 JOE, VIC, HEX, TET, CL Fluor Gold 54, Yakima Yellow Orange 585 ± 5 61 ± 5 ROX, CL Fluor Red 61, Cy 3.5, Texas Red, lexa Fluor 568 Red 625 ± 5 66 ± 1 Cy5, Quasar 67, Lightr Red64, lexa Fluor 633 Crimson 68 ± 5 712 high pass Quasar 75, Lightr Red75, lexa Fluor 68 HRM 46 ± 2 51 ± 5 SYR Green I, SYTO9, LC Green, LC Green Plus+, EvaGreen Rotor-Gene Q Rotor-Gene Q PCR Rotor-Disc Rotor-Disc Rotor-Disc 72 Rotor-Dsic 72 1 Rotor-Disc 1 Rotor-Disc Rotor- Disc Heat Sealer QIgility PCR PCR QIgility Rotor-Gene Q QIgility PCR www.qiagen.com/goto/qigility 4 Rotor-Gene Q 7/215
HRM High Resolution Melt HRM PCR PCR GC T m PCR T m High Resolution Melt HRM T m Rotor-Gene Q ±.2 PCR Rotor-Gene Q HRM 15 95 15 85 75 95 65 85 Heterozygote Homozygote 55 75 mutant 45 65 Heterozygote Homozygote mutant 35 55 25 45 35 Wild type 15 25 5 Wild type 15 78 79 8 81 82 83 84 85 86 87 5 Temperature ( C) 78 79 8 81 82 83 84 85 86 87 1 Heterozygote 5 Rotor-Gene Q HRM Heterozygote 5 HRM Wild type ±.2 78 79 8 81 82 83 Wild 84 type 85 86 87 Temperature ( C) 78 79 8 81 82 83 84 85 86 87 Temperature ( C) 3. HRM SNP PPP1R14 protein phosphatase 1, regulatory Rotor-Gene Q class IV SNP T HRM [inhibitor] subunit 14 SNP rs631276 G DN 1 ng Type-it HRM PCR 1 PCR HRM Kit Rotor-Gene Q 9 GG G 8 1% methylated HRM wt 7 difference plot 3 6 9%. 7% 5 4 4 5%. 3 3%. 2 1%. 1 % methylated 75 76 77 78 79 8 81 82 83 84 1 9 8 1% methylated 7 6 9%. 7% 5 4 5%. 3 3%. 2 1%. 1 % methylated 75 76 77 78 79 8 81 82 83 84 Temperature ( C) minus 5% meth. 3 2 1-1 -2 Temperature ( C) Normalized fluorescence minus minus wt wt 75 76 77 78 79 8 81 82 83 84 Temperature ( C) 15 15 1 Temperature ( C) Homozygote mutant Homozygote mutant 4. HRM DN-PC adenomatosis polyposis coli EpiTect HRM PCR Kit Rotor-Gene Q HRM HRM 5 difference plot minus 5% meth. 3 2 1-1 -2 75 76 77 78 79 8 81 82 83 84 Temperature ( C) Rotor-Gene Q 7/215 5
Rotor-Gene Q PCR 2 2. Rotor-Gene Q 2 C T Export to Relative Expression Software Tool REST Export to LinRegPCR assumption-free analysis Melt HRM Scatter Plot Virtual mode Rotor-Gene Q Rotor-Gene Q LED 6 Rotor-Gene Q 7/215
Rotor-Gene Q Rotor-Gene Q Rotor-Disc OTV Optical Temperature Verification Kit Rotor-Disc Rotor-Gene Q Rotor-Gene Q PCR 3 3. Rotor-Gene Q 37 mm x 42 mm x 286 mm 538 mm 12.5 kg ±.2 ±.5 ±.2 35 99 6 365 68 nm 46 75 nm LED Gain PMT 1 1 Rotor-Disc 1 3 µl x 1 15 2 µl Rotor-Disc 72.1 ml x 72 2 25 µl*.1 ml Strip Tubes.1 ml x 72 1 5 µl PCR.2 ml.2 ml x 36 2 5 µl QIGEN Rotor-Gene Kit 45 6 4 1 24 V C 5/6 Hz 56 V peak *.1 ml Strip Tubes 2 µl SN Rotor-Gene Q 7/215 7
Rotor-Gene Q QIGEN PCR 4 5 PCR Q-ond 6 45 PCR Factor MP PCR Factor MP DN 4. QIGEN DN PCR SYR Green PCR 2 RT-PCR Rotor-Gene SYR Green PCR Kit 5 SYR Green 1 RT-PCR Rotor-Gene SYR Green RT-PCR Kit 7 singleplex PCR 2 RT-PCR Rotor-Gene Probe PCR Kit 2 singleplex 1 RT-PCR Rotor-Gene Probe RT-PCR Kit 8 multiplex PCR 2 RT-PCR Rotor-Gene Multiplex PCR Kit 9 mirn SYR Green 2 RT-PCR miscript SYR Green PCR Kit singleplex PCR 2 RT-PCR QuantiTect Virus +ROX Vial Kit 1 multiplex 1 RT-PCR SNP PCR Type-it Fast SNP Probe PCR Kit 11 EvaGreen HRM Type-it HRM PCR Kit 3 PCR EpiTect MethyLight PCR Kit 12 EvaGreen HRM EpiTect HRM PCR Kit 4 NH4 + K + NH3 + H + K + P _ H P _ 5. QIGEN PCR NH 4 + K + K + DN NH 4 + Fast 6. Rotor-Gene Master Mix Type-it Fast SNP PCR Master Mix Q-ond DN DN DN Q-ond 8 Rotor-Gene Q 7/215
Rotor-Gene SYR Green Kit QuantiTect Primer ssay Rotor-Gene Q QuantiTect Primer ssay GeneGlobe www.qiagen.com/geneglobe Rotor-Gene SYR Green Kit PCR 7 5.8.6.4.2. df/dt QIGEN 1.2 1..8.6.4.2. 5 7 6 8 9 1 Temperature ( C) 5 1 15 2 25 3 35 4 45 s 5. SYR Green RT-PCR R QIGEN II C T Mean deviation C T Mean deviation X CL2-associated X protein 24.84.5 29.57.46 CL2 apoptosis gene 26.96.5 32.83.29 MYC proto-oncogene 28.42.21 35.26.72 b-ctin housekeeping gene 2.24.3 24.39.12 cdn 1 ng SYR Green 2 RT-PCR 4 QuantiTect Primer ssay triplicate X CL2 MYC b-ctin Rotor-Gene Q Rotor-Gene SYR Green RT-PCR Kit II C T TaqMan Gene Expression ssays Rotor-Gene Probe Kit Rotor-Gene Q.3 1 RT-PCR 8 QIGEN.2.1. 1 2 3 4 II 7. SYR Green RN 1 1 ng 1 pg SYR Green 1 RT-PCR CL2 -cell CLL/lymphoma 2 QuantiTect Primer ssay duplicate Rotor-Gene Q Rotor-Gene SYR Green RT-PCR Kit 1 pg RN PCR R Mg 2+ 1 pg PCR 6 E-1.3.2.1. 1 2 3 4 Rn 5 E-1 4 E-1 3 E-1 2 E-1 1 E-1 5 1 15 2 25 3 35 4 6 E-1 8. 5 E-1 RN 1 1 ng 1 ng 1 4 E-1RT-PCR IL12R1 interleukin 12 receptor, beta 1 TaqMan Gene Expression ssay3 E-1 Triplicate Rotor-Gene Q Rotor-Gene Probe RT-PCR Kit 2 E-1 II C T 1 E-1 Rn 5 1 15 2 25 3 35 4 Rotor-Gene Q 7/215 9
Rotor-Gene Multiplex PCR Kit Rotor-Gene Q 4 cdn 9 C T C TNF tumor necrosis factor IFNG interferon, gamma MYC v-myc myelocytomatosis viral oncogene homolog [avian] HSP91 heat shock protein 9 kda alpha [cytosolic], class member 1 TNF FM Cy5 HEX Texas Red CT values D 29 28 27.25 26.2 25 24.15 23.1 22.5. 8 6 4 2 TNF N.D. 1 2 3 4 TNF IFNG IFNG MYC MYC HSP9 HSP9 1 2 3 4 IFNG.25.2.15.1.5. 8 6 4 2 1 2 3 4 1 2 3 4.16.14.12.1.8.6.4.2 2 1 1 2 3 4 1 2 3 4 9..16.14 cdn 1 1 ng 1 pg 4-plex PCR Rotor-Gene 2 Q Rotor-Gene Multiplex PCR Kit S triplicate.12 1 TaqMan 4 C T S IFNG.1 N.D. Rotor-Gene Q C T C TNF S.8 D IFNG S.6 1 2 3 4.4.2 mirn 1 2 3 4 mirn QIGEN 3 miscript PCR System mirn mrn cdn SYR Green PCR mirn miscript II Reverse Transcription Kit miscript SYR Green PCR Kit miscript Primer ssay Rotor-Gene Q mirn miscript PCR System www.qiagen.com/mirn 1 Rotor-Gene Q 7/215
-2. QuantiTect Virus +ROX Vial Kit Rotor-Gene Q 4 DN RN singleplex RN DN 1 QIGEN II 1 -.2 1 -.5 1 -.8 1-1.1 1-1.4 1-1.7 1-1. 1-1.2 1-1.4 1-1.6 1-1.8 1 1 2 3 4 5 1 2 3 4 5 1..16 5 influenza virus RN1 4 in vitro duplex.14 RT-PCR singleplex 1 RT-PCR TaqMan.12 triplicate Rotor-Gene Q QuantiTect Virus +ROX Vial Kit duplex.1 singleplex.8 II duplex.6 singleplex.4.2 5 1 15 2 25 3 35 4 SNP Type-it Fast SNP Probe PCR Kit SNPs Rotor-Gene Q SNP 11 SNP TaqMan MG TaqMan 11. SNP Type-it Fast SNP Probe PCR Kit 7 DN 1 ng llelic discrimination rs 951134 TaqMan SNP 2 NTCs Rotor-Gene Q PCR NTCs FM T allele VIC allele Rotor-Gene Q 7/215 11
DN HRM 5 4 CpG EpiTect MethyLight PCR Kit Rotor-Gene Q TaqMan EpiTect MethyLight ssay EpiTect MethyLight ssay PCR 2 single PCR 12 excitation excitation C Primer 5' 3'.6.5.4.3.2.1. excitation FM FM FM 5 15 25 35 VIC Q Q excitation Primer.4 5'.3 3'.2 3' 5' Primer.1 Fluorophore 1 (FM). Fluorophore 2 (VIC) Quencher VIC Q 5 15 25 35 D VIC.16 1%.14 9%.12 45 5% 1% % NTC.6.5.1.8.6.4.2. VIC 5 1 15 2 25 3 35 4 FM Q 1% 9% 5% 3' 5' Primer % NTC 45 1% % 1% 5% 9% 1% NTC 45 Q.16 C T CG FM.14 CG FM C T TG % VIC TG VIC 1% threshold C meth = 1/ 1 + 2 C T CG C T TG.12threshold 5%.1.8 9%.6 C.4 T CG C T TG FM VIC.2 1% NTC. 45 3.37.4 1 5 1 15 2 25 3 35 4 45 34.62 3.72 6.278 5 31.27 31.67 56.887 9 3.52 33.43 88.258 1 3.5 45 99.997 12. EpiTect MethyLight PCR Kit DN 2 TaqMan PCR isulfite DN FM TaqMan isulfite DN VIC TaqMan DN PCR 1 ng isulfite DN DN 9 1 DN Hs_TMEFF2 EpiTect MethyLight ssay EpiTect MethyLight PCR Kit Rotor-Gene Q DN C T CG FM C T C DN C T TG VIC C T D 12 Rotor-Gene Q 7/215
Cat. no. Rotor-Gene Q 2plex System* Rotor-Gene Q 2plex HRM System* Rotor-Gene Q 5plex System* Rotor-Gene Q 5plex HRM System* Rotor-Gene Q 6plex System* Rotor-Disc 1 Starter Kit Rotor-Gene Q 2plex green yellow Rotor-Gene Q Software PC Rotor-Gene Q 2plexHRM green yellow HRM Rotor-Gene Q Software PC Rotor-Gene Q 5plex green yellow orange red crimson Rotor-Gene Q Software PC Rotor-Gene Q 5plex HRM green yellow orange red crimson HRM Rotor-Gene Q Software PC Rotor-Gene Q 6plex blue green yellow orange red crimson Rotor-Gene Q Software PC Rotor-Disc 1, Rotor-Disc Heat Sealer, Rotor-Disc Heat Sealing Film, Rotor-Disc 1 Rotor and Locking Ring, Rotor-Disc 1 Loading lock, Rotor-Disc Pipetting id 9162 2,5, 9163 3,9, 9164 3,5, 9165 4,9, 9166 4,2, 9162 69, Rotor-Disc 1 (3) Pack of 3 individually wrapped discs for 3 reactions 981311 33,5 Rotor-Disc 72 Starter Kit Rotor-Disc 72, Rotor-Disc Heat Sealer, Rotor-Disc Heat Sealing Film, Rotor-Disc 72 Rotor and Locking Ring, Rotor-Disc 72 Loading lock, Rotor-Disc Pipetting id 916 66, Rotor-Disc 72 (24) Pack of 24 individually wrapped discs for 1728 reactions 98131 29,5 Strip Tubes and Caps,.1 ml (25) PCR Tubes,.2 ml (1) 25 strips of 4 tubes and caps for 1 reactions 98113 22, 1 thin-walled tubes for 1 reactions 9815 1, Rotor-Disc OTV Kit Kit for optical temperature verification of Rotor-Gene Q systems; requires Rotor-Disc 72 Rotor and Locking Ring or Rotor-Disc 72 Starter Kit 9814 46, Rotor-Gene SYR Green For 4 x 25 µl reactions: 2x Master Mix, RNase-Free Water 2474 5, PCR Kit (4) Rotor-Gene SYR Green RT-PCR Kit (4) For 4 x 25 µl reactions: 2x Master Mix, RT Mix, RNase-Free Water 24174 1, Rotor-Gene Probe PCR For 4 x 25 µl reactions: 2x Master Mix, RNase-Free Water 24374 55,5 Kit (4) Rotor-Gene Probe RT-PCR Kit (4) For 4 x 25 µl reactions: 2x Master Mix, RT Mix, RNase-Free Water 24574 79, Rotor-Gene Multiplex PCR For 4 x 25 µl reactions: 2x Master Mix, RNase-Free Water 24774 72,5 Kit (4) * Rotor-Gene Q 7/215 13
Cat. no. QuantiTect Primer ssay (2)* For 2 x 5 µl reactions: 1x QuantiTect Primer ssay (lyophilized) Varies 18, QuantiTect Reverse Transcription Kit (5)* miscript SYR Green PCR Kit (2)* QuantiTect Virus +ROX Vial Kit (2)* For 5 x 2 µl reactions: uffers, Quantiscript Reverse Transcriptase, RT Primer Mix, RNase-Free Water For 2 x 5 µl reactions: 2x Master Mix, Universal Primer, RNase-Free Water For 2 x 5 µl reactions: 5x Master Mix, RT Mix, RNase-Free Water, Nucleic cid Dilution uffer 25311 39, 21873 53,5 21133 15, Type-it Fast SNP Probe PCR Kit (8)* EpiTect MethyLight PCR Kit (2)* For 8 x 25 µl reactions: 2x Master Mix, RNase-Free Water, 2645 128, Q-Solution For 2 x 5 µl reactions: 2x Master Mix, RNase-Free Water 59436 74,5 EpiTect HRM PCR Kit (1) For 1 x 25 µl reactions: 2x Master Mix, RNase-Free Water 59445 18,5 Type-it HRM PCR Kit (1) For 1 x 25 µl reactions: 2x Master Mix, RNase-Free Water 26542 19, * miscript PCR System www.qiagen.com/mirn Rotor-Gene Q QIGEN Kit QIGEN Kit Rotor-Gene Q QIGEN www.qiagen.com Trademarks and Disclaimers QIGEN Handbook User Manual www.qiagen.com Rotor-Gene Q www.qiagen.com/goto/rotor-geneq 14 Rotor-Gene Q 7/215
Trademarks: QIGEN, EpiTect, GeneGlobe, Q-ond, Q-Solution, Quantiscript, QuantiTect, Rotor-Gene, Rotor-Disc, Type-it (QIGEN Group); lexa Fluor, SYR, Texas Red (Molecular Probes, Inc.); CL Fluor, Quasar (iosearch Technologies, Inc.); Cy (GE Healthcare); FM, JOE, ROX, VIC (pplera Corporation or its subsidiaries); Lightr, TaqMan (Roche Group). 215 QIGEN, all rights reserved. Rotor-Gene Q 7/215 15
www.qiagen.com 14-54 3-13-1 Forefront Tower II Tel:3-689-73 Fax:3-5547-818 E-mail:techservice-jp@qiagen.com 232299 7/215