TaKaRa PCR FLT3/ITD Mutation Detection Set
Contents
1. Agilent2100 Agilent Technologies LabChip Caliper Technologies Corporation TaKaRa PCR FLT3 ITD Mutation Detection Set TEL 077 543 7231 FAX 077 543 9254 NOTICE TO PURCHASER LIMITED LICENSE M31 FLT3 ITD mutation detection This product is cover2. Control Template Normal ITD ou ui IO http www takara bio co jp 4 6632 V AML DNA Control Template Mutant Control Template Normal IIV PCR PCR 3 NuSieve 3 1 Control Template Normal 329 bp Control Template Mutant 359 bp AML DNA 329 bp 60 bp ITD M 1 2 8 359 bp 400 bp 329 bp 300 bp gt 1 AML DNA Control Template M 100 bp DNA Ladder Marker 1 AML DNA 2 Control Template Mutant 359 bp 3 Control Template Normal 329 bp
3. PCR 2 3 7 8 9 10 B PCR 1 2 1 ml 5 ml 1500 rpmy 5 Hemolysis Buffer 15 ml 1500 rpm 10 1 Hemolysis Buffer 10 mM Tris HCI pH7 6 5 mM MgCl2 10 mM NaCI 2 2 Lysis Buffer 21 5 ml 37 2 2 Lysis Buffer 10 mM Tris HCI pH7 6 10 mM EDTA 50 mM NaCl 0 5 SDS 0 1 mg ml proteinase K 25 24 1 12000 rpm 5 24 1 12000 rpm 5 5 M NaCI 30 ul 0 9 ml 12000 rpm 5 70 TE Buffer
4. Migration time Fluorescence N Control Template Normal M Control Template Mutant http www takara bio coJp 6 6632 ITD 80 70 60 50 30 Fluorescence 20 B 85 45 6 DOL GOL OLL 0ZL GL 1 Cm Dr gy 09 e9 09 s9 w Se 0 Time seconds 3 AML DNA Agilent2100 N ITD ITD VI 1 Nakao M Yokota S Iwai T et al 1996 Leukemia 10 1911 1918 2 Yokota S Nakao M Okuda T et g 1997 Leukemia 11 1605 1609 3 Kiyoi H Naoe T Yokota S et al 1997 Leukemia 11 1447 1452 4 Kiyoi H Naoe T Nakao Y et al 2002 Blood 93 3074 3080 5 Kiyoi H Naoe T 2002 Leukemia and Lymphoma 43 1541 1547 6 Zheng R Friendman AD Small D 2002 Blood 100 4154 4161 http www takara bio coJp 7 6632
5. http www takara bio cojp 5 6632 B Agilent2100 Control Template Mutant Control Template Normal AML DNA IV PCR DNA 1000 LabChip 2100 Control Template Mutant Control Template Normal 329 bp Normal 359 bp Mutant 2 2 AML DNA 329 bp 60 bp ITD 3 N LM 8 GC 8 Me 3 1 an Ri _ e Eee ig Eg ow nn NN oo em ee Cn oo oo cn o ow wn 1 4 Time seconds 2 Control Template Agilent2100
6. F6032 IdfaR4 TaKaRa PCR FLT3 ITD Mutation Detection Set vV201210Da FLT3 FMS like tyrosine kinase 3 JM Juxtamembrane Inter nal Tandem Duplication ITD FLT3 ITD AML 1 3 MDS 3 JM exon11 intron11 exon12 exon11 exon12 ITD DNA PCR Agilent2100 DNA 5 10 ITD
7. DNA 0 2 ml PCR tube 1 10 Xx Ex Tag Buffer Mg plus 1 5 ul dNTP Mixture 2 5 mM 1 4 ul FLT3 ITD 1 Primer 10 pmol ul 1 ul FLT3 ITD 2 Primer 10 pmol ul 1 ul TakaRa Ex 7g9 5 U ul 0 25 wl DNA 2 0 1 ug up to 50 wl 1 7aKaRa Ex Tag 2 Control Template 1 ul PCR 94 30 sec 56 30 sec 35 cycles 72 30 sec 72 5 min http wwwr takara bio coJp 3 6632 C PCR 5 3 NuSieve 3 1 Agarose TAE 100 bp DNA Ladder UV Control Template Normal Mutant 329 bp ITD
8. 329 bp 347 419 bp ITD 3 Control Template Normal 329 bp Control Template Mutant 359 bp 3 ITD 329 bp ITD 347 419 bp 2 ITD D Agilent2100 Agilent2100 PCR TE buffer 0 5 50 ng ul 2 10 DNA1000 LabChip Gel Dye Mix LabChip Gel Dye Mix LabChip LabChip 1 ul LabChip 1 LabChip DNA 1000 12 30
9. AML FLT3 1 50 1 FLT3 ITD 1 Primer 10 pmoluD 50 ul 50 2 FLT3 ITD 2 Primer 10 pmol ul 50 ul 50 3 Control Template Mutant 1 pg uD 10 10 4 Control Template Normal 1pg uD 10 10 20C 7aKaRa Ex Tag RR001A NuSieve 3 1 Agarose 50091 buffer TAE buffer DNA 100 bp DNA ladder 3407A B Loading buffer 6 X 36 glycerol 0 05 bromophenol blue 0 05 xylene cyanol 30 mM EDTA DNA DNA SYBR Green 1 50512 50513 GelStar Nucleic Acid Stain 50535 authorized instruments TaKaRa PCR Thermal Cycler Dice
10. Gradient Standard TP600 TP650 TaKaRa PCR Thermal Cycler Dice mini TP100 Mupid 2plus M 2P Mupid exU EXU 1 UV 300 nm SYBR Green GelStar Nucleic Acid Stain 0 2 ml PCR tube 0 2 ml Hi Tube Dome Cap NJ200 0 5 mL1 5 ml http wwwr takara bio coJp 2 6632 Agilent2100 IV Agilent2100 Agilent DNA1000 LabChip A DNA DNA PCR EDTA 2Na DNA
11. ed by the claims of U S Patent No 6 846 630 and its foreign counterpart patent claims TakaRa Tel 077 543 6116 Fax 077 543 197 http wWW taKara DiO CO jp v201210Da
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