DNA. Application Note. DNA dsdna DNAPac RP HPLC. DNAPac RP. dsdna PCR DNA. dsdna. CRISPR-Cas9 DNA DNA. SNPs. DNA dsdna. dsdna HPLC

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Application Note DNA 21533 DNA dsdna DNAPac RP HPLC DNAPac RP dsdna PCR DNA dsdna CRISPR-Cas9 DNA 1 SNPs

1 Application Note DNA DNA dsdna DNAPac RP HPLC DNAPac RP dsdna PCR DNA dsdna CRISPR-Cas9 DNA 1 SNPs 2 DNA dsdna dsdna DNA HPLC DNA dsdna Thermo Scientific DNAPac RP4 m ph 1 2 Å dsdna DNAPac RP DNA DNA PCR

2 18.2 McmDI Fisher Scientific P/N A955-4 TEAA 2. M Applied Biosystems P/N 4613 pbr322-bsurithermo Scientific P/N SM252 ΦX174-BsuRIThermo Scientific P/N SM271 Thermo Scientific FastRuler Low Range DNA Ladder ready-to-use P/N SM113 Thermo Scientific FastRuler Middle Range DNA Ladder ready-to-use P/N SM1113 Thermo Scientific FastRuler High Range DNA Ladder, readyto-use P/N SM1123 WPRE PCR Sanjay Vasu P/N LP/N TCC-3RS P/N 573. P/N VWD-34RS UV/VisP/N DNAPac RP mm P/N DNAPac RP mm P/N A.1 M TEAA ph 7. B.1 M TEAA /75:25 v/v UV26 nm 3 L PP P/N C413-11PTFE PP P/N C413-77A Thermo Scientific Invitrogen 1.2% E-Gel P/N G5181 Invitrogen E-Gel Base FisherBiotech FB2Q UV/Vis Thermo Scientific Dionex Chromeleon CDS 7.2 pbr322-bsuriφx174-bsuri 5 1 g/mlfastruler DNA Ladder PCR 1.Thermo Scientific Vanquish Flex UHPLC P/N VF-S1-A Flex P/N VF-P2-A FT P/N VF-A1-A H P/N VH-C1-A P/N HL P/N VH-D1-A Thermo Scientific LightPipe 1 mm P/N Thermo Scientific Dionex UltiMate 3 BioRSLC SR-3 P/N LPG-34RS RS P/N WPS-3TBFC 2

3 DNAPac RP DNA 1BsuRIpBR322 1a ΦX174DNA1bdsDNA 55 bp 1 DNAPac RP DNA 1cpBR322ΦX a Absorbance mau (mau) 8 b Retention Time (min) Retention Time (min) c pbr322-bsuri digest X 174-BsuRI digest y = -4.52x² x R² =.999 DNAPac RP 4 m mm A.1 M TEAA ph 7. B.1 M TEAA ph 7. /75:25 v/v A B ml/min 5 L 55 UV 26 nm a. pbr322-bsuri1 g/ml b.φx174-bsuri1 g/ml Log (number of base pairs) 1 3

4 DNAPac RP dsdna 1, DNA 2DNA DNA 14 1, bp1 DNA DNADNA 28 a k 5 mau Absorbance (mau) 28 b k 5k 28 c 5 1k 4k 1k 2k Retention Time (min) DNAPac RP 4 m mm A.1 M TEAA ph 7. B.1 M TEAA ph 7. /75:25 v/v A B ml/min 1 L 55 UV 26 nm DNA a. b. c. 2 FastRuler DNA Low range bp Middle range bp High range 2% 2 agarose 1 1% agarose 1% 1 agarose bp bpφx174-bsuri DNAPac RP 2.1 mm IDC 2. mm ID DNAPac RP dsdna 3 4

5 mau Absorbance (mau) DNAPac RP, 1~2 Å, 4 μm Brand A 3 Å, 1.7 μm Brand B 1 Å, 2.7 μm 35 Brand C 11 Å, 3 μm Retention Time (min) 3 ΦX174-BsuRI ΦX174-BsuRI dsdna 4 dsdna5 6 6 dsdna 65 DNA ssdna dsdna ssdna 8 9 dsdna mau Absorbance (mau) ºC 4 ºC 5 ºC 6 ºC 7 ºC DNAPac RP 4 m mm A.1 M TEAA ph 7. B.1 M TEAA ph 7. /75:25 v/v A B ml/min 5 L UV 26 nm ΦX174-BsuRI1 g/ml ep 14 8 ºC 1 9 ºC Retention Time (min) 4ΦX174-BsuRI 5

DNAPCR PCRPCRdNTP UltiMate 3 UltiMate WPS-3TBFC PCR 5 4,5 1.2E-Gel 5bDNAPac RP HPLC dsdnapcr 2 a 4 3 mau Absorbance (mau) 2 1 dntp impurities PCR product DNAPac RP 4 m 2.1 1 mm A.1 M TEAA ph 7. B.

6 DNAPCR PCRPCRdNTP UltiMate 3 UltiMate WPS-3TBFC PCR 5 4,5 1.2E-Gel 5bDNAPac RP HPLC dsdnapcr 2 a 4 3 mau Absorbance (mau) 2 1 dntp impurities PCR product DNAPac RP 4 m mm A.1 M TEAA ph 7. B.1 M TEAA/75:25 v/v A B L 5 UV 26 nm PCR Retention Time (min) b PCR 5 PCR DNAPac RPVanquish Flex UHPLC 1, bp dsdna dsdna bp WPS-3 TBFC UltiMate 3 dsdna 6

7 1. Sander, J.D.; Joung, J.K. CRI SPR-Cas systems for editing, regulating and targeting genomes, Nature Biotechnology, 214, 32, Wolff, J.N.; Gemmell, N.J. Combining allele-specific fluorescent probes and restriction assay in real-time PCR to achieve SNP scoring beyond allele ratios of 1:1, BioTechniques, 28, 44, Baek, J.; Thayer, J.; Lin, S.; Liu, X. Separation of Mixed-Base Oligonucleotides Using a High-Resolution, Reversed-Phase Chromatography Column, Thermo Fisher Scientific AN (216). 4. Thayer, J.R., Puri, N., Burnett, C., Hail, M. and S. Rao. Identification of linkage isomers by anion exchange purification with electrospray ionization mass spectrometry of automatically desalted phosphodiesterase-ii digests. Analytical Biochemistry, 21, 399, Thayer, J.R., Flook, K.J., Woodruff, A., Rao, S., Pohl, C. New monolith technology for automated anion-exchange purification of nucleic acids. Journal of Chromatography B, 21, 878, 933. AppsLab Library eworkflow Chromeleon cmbx 7

8 216 Thermo Fisher Scientific Inc. HPLC57_A171SO TEL FAX

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