DNA Damage Quantification Kit
Contents
1. 4 DNA 5 630 670 nm 50 ll 1 mol l 450 nm 6 A 1 DNA DNA Binding Solution DNABinding Solution DNA 14 000 HRP Streptavidin Substrate Solution2. 650 nm DNA AP site d 650 nm 50 pl 50 1 mol 450 nm 2025 5 861 2202 Tel 096 286 1515 Fax 096 286 1525 URL www dojindo coJp Tel 03 3578 9651 Fax 03 3578 9650 K02 Nucleostain DNA Damage Quantification Kit AP site Counting kit
3. DNA DNA polymerase DNA AP site apurinic apyrimidinic site AP site DNA Nucleostain DNA Damage Quantification Kit AP Site Counting AP site ARP V Amino oxymethylcarbonyI hydrazino D biotin DNA 96 DNA AP site DNA AP site ARP Standard DNA DNA AP site
4. 400 pl ARP Buffer ARP DNA ARP DNA ARP DNA Filration Tube ee 2500xg DNA 1 5ml 2500 x g 15 5 sige 2OHI ARP DNA 310 MM TE Buffer 60 ul ARP DNA Standard Solution ARP DNA 0 40 ARP DNA Standard Solution b 2 Se 29 100 Ll DNA Binding Solution Ez overnight K02 Nucleostain DNA Damage Quantification Kit AP site Counting kit General Protocol at a Glance General Protocol ee c 1 4000 HRP Streptavidin 10 pl HRP Streptavidin 40 ml PBST
5. ARP DNA 0 5C DNA TE Buffer TE Buffer 500 ml 500 ml Tris 606 mg 10 mmoMl 3NA 206 mg 1 mmol l 6 moll HCI pH 7 5 2 500 x 9 5 Filtration Tube DNA 90 ARP DNA 2 25 ug ml 2 AP site 1 1 ARP DNA 90 ul 1 5 ml TE Buffer 310 2 ARP DNA Standard Solution 1 60 ul 3 2 3 1 ARP DNA 1 60 pl 3 4 DNA Binding Solution 100 pl DNA 5 Washing Buffer 1L PBST
6. PBST PBST Washing Buffer 1L PBST DNA DNA 4 6 ARP Filtration Tube ARP 7 1 1 Filration Tube 1 E DNA 1 DNA 0ARP DNA Standard Solution DNA AP site 0 ARP DNA Standard Solution
7. HRP Streptavidin PBST 4 000 HRP Streptavidin HRP Streptavidin 10 pl PBST 40 ml 6 PBST 250 ul 5 7 HRP Streptavidin 150 ul 37 C 1 8 6 PBST 5 9 Substrate Solution 100 pl 37 C 1 10 630 670 nm 11 DNA AP site 3 ARP DNA Standard Solution ARP DNA 60 DNA Binding Solution 100 5 samples 20 samples 1 2 3 4 5 6 7 8 9 10 11 12 1 2 9 6 7 9 9 1 0 ARP DNA std 25ARPDNAsd 5 EE SARP DNA std use these ea fo sample DNA sample 5 sample 6 sample 7 sam
8. 2 Filration Tube DNA TE Buffer 3 DNA T Lindahl and B Nyberg Biochemistry 1972 11 3610 J Nakamura V E Walker P B Upton S Y Chiang Y W Kow and J A Swenberg Cancer Res 1998 58 222 K Kubo H lde S S Wallace and Y W Kow Biochemistry 1992 31 3703 H Ide K Akamatsu Y Kimura K Michiue K Makino A Asaeda Y Takamori and K Kubo Biochemistry 1993 32 8276 A Asaeda H lde K Tano Y Takamori and K Kubo Nucleosides amp Nucleotides 1998 17 503 nwon General Protocol at a Glance General Protocol Day 1 Sle OU DNA 10 ARP Solution 37 1 a DNA 10 ARP Solution 400 pl ARP DNA DNA concentration Step 1 ARP DNA 20 pl 380 pl spin Sn 2 25 ngml TE Buffer 17 frate Step Step ii Step iii cup 100 TEBuffer 2 400l TE 200ul TEBuffer
9. 20 samples 250 pl x 1 10 ml x 1 10 ml x 1 40 ml x 1 25 IX 1 1 packet 20 tubes EE TT 20 samples 250 ul x 1 250 ul x 1 10 ml 200 1 ml ARP DNA Standard Solution 0 ARP DNA Standard Soln 0 ARP 100 000 bp 2 5 ARP DNA Standard Soln 2 5 ARP 100 000 bp 5 ARP DNA Standard Soln 5 ARP 100 000 bp 10 ARP DNA Standard Soln 10 ARP 100 000 bp 20 ARP DNA Standard Soln 20 ARP 100 000 bp 40 ARP DNA Standard Soln 40 ARP 100 000 bp 0 5 ml 1 5 ml 200 ul 8 37 C DNA DNA AP site RNA RNase A RNA DNA DNA DNA DNA
10. Get pureDNA Kit Agarose Get pureDNA Kit Cell Tissue DNA 0120 489548 400 ul 200 ulx2 7 il Rt ARP DN 2 500 x g 15 CN 2 500 x g 15 ARP DNA SteD 2 37C 1 TE Buffer 380 Hl DNA i 10Ll ARP solution 10 Hl 400 ul 2 25 ug ml Step 1 cup 100 TE Buffer 2 400 ll ARP DNA Filtration Tube 2 500 x g 15 400 pl TE Buffer Filtration Tube 2 500 x g 15 1 DNA ARP Step 3 200 ul TE Buffer DNA 1 5 ml 200 ul TE Buffer DNA 1 5 ml 1 DNA ARP 1 1 DNA TE Buffer 260 nm DNA 5
11. HRP Streptavidin HRP Streptavidin 0 5C DNA DNA 100 ugml DNA DNA ARP Solution ARP Filtratlon Tube DNA AP site 17100 000 bp C 1 HRP Streptavidin PBST 14 000 HRP Streptavidin PBST 5 RwoMN Dm Dm nr ow nm 9 ARP DNA Standard SoIn PBST 5 PBST
12. 0 ug ml 1 0 100 ug ml TE Buffer 2 DNA 10 ul ARP Solution 10 pl 0 5 ml 3 37 C 1 380 ul TE Buffer 4 Filtration Tube cup 100 l TE Buffer 2 3 Filtration Tube 5 2 500 x g 15 6 5 Filtration Tube 400 ul TE Buffer ARP DNA 3 7 2 500 x 9 15 8 7 Filtration Tube 200 ul TE Buffer ARP DNA 3 9 DNA 1 5 ml 10 200 ul TE Buffer 8 Filtration Tube 8 ARP DNA 8 DNA 11 1 5 ml
13. ple 8 sample 9 10ARPDNAsd 20ARPDNAsd 40 ARP DNA std blank 60 pl TE buffer 100 pl DNA binding solution well sample 10 sample 1 MT OO OO Ww TT MT OU ODO Ww sample 12 250 ul well x5 250 ul well x5 HRP Streptavidin 150 ul well HRP Streptavidin 150 ul well 250 ul well x5 250 ul well x5 Substrate Solution 100 pl well Substrate Solution 100 ul well Washing Washing HRPistreptavidin i i Ne eSubgtrate sotutign Sadss Sea ess RR E RN a HRP streptavidin TT Mm OU ODO Ww D gt TT Mm OU OO Ww 2 0 8 0 6 0 4 Absorbance at 650 nm 0 2 0 10 20 30 40 Number of AP sites 100 000 bp 3 ARP DNA Standard Solution 1 0 lt 5C 2 DNA AP site DNA ARP ARP DNA 0 5C 1 3 Filtration Tube TE Buffer DNA DNA DNA
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