1 buffer 176 enzymes 100% activity - Carlova Consult...

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t # M

B145

8

Single letter code B = C, G or T K = G or T R = G or A W = A or TD = A, G or T M = A or C S = C or G Y = C or TH = A, C or T N = G, A, T or C V = A, C or G

Cleavage blocked or impaired by Dam methylation within the recognition sequenceCleavage blocked or impaired by Dcm methylation within the recognition sequenceCleavage blocked or impaired by CpG methylation within the recognition sequenceOverlapping Dam methylation may block or impair DNA cleavageOverlapping Dcm methylation may block or impair DNA cleavageOverlapping CpG methylation may block or impair DNA cleavage

Methylation sensitivity

For detailed information, see Certifi cate of Analysis supplied with each enzyme.

Activity of DNA/RNA modifying enzymes inFastDigest Green Buffer and FastDigest Buffer

DNA/RNA modifying enzyme Activity in FastDigest Green Buffer/ FastDigest Buffer

DNA Polymerase I (#EP0041) 100%Klenow Fragment (#EP0051) 100%Klenow Fragment, exo– (#EP0421) 100%T4 DNA Polymerase (#EP0061) 100%T7 DNA Polymerase (#EP0081) 100%T4 DNA Ligase* (#EL0011) 75-100%Thermo Scientifi c™ FastAP™ Alkaline Phosphatase (#EF0651) 100%

T4 Polynucleotide Kinase (#EK0031) 100%

*0.5 mM ATP is required for T4 DNA Ligase activity.

Standard reaction setup

Component Plasmid DNA

PCR product Genomic DNAunpurifi ed purifi ed

Nuclease-free water 15 µL 17 µL up to 30 µL 30 µL10X FastDigest Green Buffer or 10X FastDigest Buffer

2 µL 2 µL 3 µL 5 µL

DNA 2 µL (up to 1 µg)

10 µL (~0.2 µg) ~0.2 μg 10 µL

(5 µg)FastDigest enzyme 1 µL 1 µL 1 µL 5 µLTotal volume 20 µL 30 µL 30 µL 50 µL

The volume of DNA can be scaled up to 10 µL or down to 0.5 µL depending on the DNA concentration.The volume of water should be corrected to keep the indicated total reaction volume.

• 100% activity of all Thermo Scientifi c™ FastDigest™ enzymes in one buffer

• Complete digestion in 5-15 minutes

• 100% buffer compatibility with downstream applications

• Direct loading on gels

• 176 FastDigest enzymes available

Five minute single, double and triple digestions in FastDigest Green Buffer

M 1 2 3 4 M

1: Plasmid - undigested control2: Plasmid - digested with FastDigest EcoRI3: Plasmid - double digested with FastDigest EcoRI and FastDigest KpnI4: Plasmid - triple digested with FastDigest EcoRI, FastDigest KpnI and FastDigest SmaIM: Marker - Thermo Scientifi c™ GeneRuler™ Express Ladder (#SM1553)

1 buffer • 176 enzymes • 100% activityThermo Scientifi c FastDigest Restriction Enzymes

Enzyme Specifi city 5’→ 3’

Reaction temperature,

°C

Digestion time with 1 µL of FastDigest enzyme, minbp from end of DNA

required for complete digestion

Thermal inactivation

Incubation time without star

activity,hours

Methylation sensitivity Cat. No.

Package size,

reactionsLambda DNA Plasmid DNA PCR product Genomic DNA

1 µg/20 µL 1 µg/20 µL ~0.2 µg/30 µL 1 µg/10 µL

FastDigest AanI TTA↓TAA 37 5 5 5 5 5 65°C, 5 min 16 no effect FD2064 20FastDigest AasI GACNNNN↓NNGTC 37 5 5 5 5 1 80°C, 10 min 1 FD1724 25FastDigest AatII GACGT↓C 37 15 20 15 15 5 80°C, 5 min 16 FD0994 50FastDigest Acc65I G↓GTACC 37 5 5 5 5 3 65°C, 5 min 16 FD0904 100FastDigest AdeI CACNNN↓GTG 37 5 5 5 5 2 80°C, 5 min 1 FD1234 50FastDigest AjuI* ↓(7/12)GAA(N)7TTGG(11/6)↓ 37 5 5 10 5 ND 65°C, 5 min 16 no effect FD1954 20FastDigest AluI AG↓CT 37 15 15 15 15 4 65°C, 5 min 16 no effect FD0014 100FastDigest Alw21I GWGCW↓C 37 5 5 5 5 1 80°C, 20 min 16 no effect FD0024 100FastDigest Alw26I GTCTC(1/5)↓ 37 5 5 5 5 1 65°C, 5 min 16 FD0034 100FastDigest Alw44I G↓TGCAC 37 5 60 10 5 4 80°C, 5 min 16 FD0044 200FastDigest ApaI GGGCC↓C 37 5 5 20 10 2 65°C, 5 min 16 FD1414 300FastDigest BamHI G↓GATCC 37 5 5 5 5 2 80°C, 5 min 1 no effect FD0054/5 800/2500FastDigest BclI T↓GATCA 37 5 5 15 5 3 80°C, 20 min 16 FD0724 300FastDigest BcnI CC↓SGG 37 5 5 5 5 3 80°C, 20 min 16 FD0064 200FastDigest BcuI A↓CTAGT 37 no sites 5 5 5 1 No 16 no effect FD1254 50FastDigest BfmI C↓TRYAG 37 5 5 5 5 2 65°C, 20 min 16 no effect FD1164 20FastDigest BfoI RGCGC↓Y 37 5 5 5 5 2 65°C, 10 min 1 FD2184 200FastDigest BglI GCCNNNN↓NGGC 37 5 5 5 5 3 65°C, 5 min 2 FD0074 200FastDigest BglII A↓GATCT 37 5 20 30 20 3 No 16 no effect FD0084 200FastDigest Bme1390I CC↓NGG 37 5 5 5 5 4 No 16 FD1424 100FastDigest BmsI GCATC(5/9)↓ 37 5 5 5 5 1 65°C, 5 min 16 FD2124 20FastDigest BoxI GACNN↓NNGTC 37 5 5 5 5 3 80°C, 20 min 16 FD1434 100FastDigest BpiI GAAGAC(2/6)↓ 37 5 5 5 5 1 65°C, 10 min 16 no effect FD1014 20FastDigest BplI* ↓(8/13)GAG(N)5CTC(13/8)↓ 37 15 20 30 30 ND 65°C, 5 min 16 no effect FD1314 20FastDigest Bpu10I CCTNAGC(-5/-2)↓ 37 15 15 30 15 3 80°C, 5 min 1 no effect FD1184 20FastDigest Bpu1102I GC↓TNAGC 37 5 5 5 5 2 80°C, 5 min 16 no effect FD0094 50FastDigest BseDI C↓CNNGG 37 5 5 5 5 3 80°C, 5 min 16 no effect FD1084 20FastDigest BseGI GGATG(2/0)↓ 37 5 5 5 5 2 80°C, 5 min 16 no effect FD0874 100FastDigest BseJI GATNN↓NNATC 65 5 5 5 5 3 No 1 FD1714 200FastDigest BseLI CCNNNNN↓NNGG 37 5 5 5 5 2 No 16 FD1204 100FastDigest BseMI GCAATG(2/0)↓ 55 15 15 15 15 3 80°C, 5 min 16 no effect FD1264 20FastDigest BseMII CTCAG(10/8)↓ 55 15 30 15 15 2 80°C, 5 min 16 no effect FD1404 20FastDigest BseNI ACTGG(1/-1)↓ 65 5 5 5 5 2 80°C, 5 min 16 no effect FD0884 100FastDigest BseSI GKGCM↓C 37 5 5 5 30 3 80°C, 15 min 16 no effect FD1444 50FastDigest BseXI GCAGC(8/12)↓ 65 15 15 15 15 3 80°C, 15 min 16 no effect FD1454 20FastDigest Bsh1236I CG↓CG 37 5 5 5 5 1 80°C, 10 min 16 FD0924 100FastDigest Bsh1285I CGRY↓CG 37 15 15 15 15 3 80°C, 15 min 1 FD0894 100FastDigest BshNI G↓GYRCC 37 5 60 5 15 2 65°C, 10 min 16 FD1004 300FastDigest BshTI A↓CCGGT 37 5 5 5 5 3 80°C, 5 min 16 FD1464 20FastDigest Bsp119I TT↓CGAA 37 5 5 5 5 1 80°C, 5 min 16 FD0124 200FastDigest Bsp120I G↓GGCCC 37 5 5 15 5 3 80°C, 10 min 16 FD0134 200FastDigest Bsp1407I T↓GTACA 37 5 5 5 5 3 80°C, 10 min 16 no effect FD0934 100FastDigest Bsp143I ↓GATC 37 5 5 10 5 4 65°C, 20 min 16 FD0784 40FastDigest BspLI GGN↓NCC 37 5 5 5 5 2 65°C, 20 min 16 FD1154 20FastDigest BspOI GCTAG↓C 37 5 5 15 5 3 80°C, 5 min 1 no effect FD2044 20FastDigest BspTI C↓TTAAG 37 5 5 5 5 4 No 16 no effect FD0834 150FastDigest Bst1107I GTA↓TAC 37 5 5 5 10 3 No 6 FD0704 100FastDigest BstXI CCANNNNN↓NTGG 37 5 5 5 5 4 80°C, 5 min 0.5 FD1024 100FastDigest Bsu15I AT↓CGAT 37 5 5 5 5 2 65°C, 15 min 16 FD0144 100FastDigest BsuRI GG↓CC 37 5 5 5 5 3 No 16 no effect FD0154 400FastDigest BveI ACCTGC(4/8)↓ 37 15 15 60 20 5 65°C, 5 min 16 FD1744 50FastDigest CaiI CAGNNN↓CTG 37 5 5 5 5 4 65°C, 10 min 16 FD1394 50FastDigest Cfr10I R↓CCGGY 37 5 5 10 5 2 No 2 FD0184 50FastDigest Cfr13I G↓GNCC 37 5 5 5 5 2 No 16 FD0194 100FastDigest CpoI CG↓GWCCG 37 5 5 5 5 1 65°C, 5 min 16 FD0744 20FastDigest CseI GACGC(5/10)↓ 37 15 15 30 15 2 80°C, 5 min 16 FD1904 20FastDigest CsiI A↓CCWGGT 37 5 10 5 15 4 65°C, 5 min 16 no effect FD2114 50FastDigest Csp6I G↓TAC 37 5 5 5 5 2 80°C, 10 min 16 no effect FD0214 100

FastDigest DpnI Gm6A↓TC 37 ND 5 not applicable 10 1 80°C, 5 min 16 does not cut dam– DNA FD1704 100

FastDigest DraI TTT↓AAA 37 5 5 5 5 4 65°C, 5 min 16 no effect FD0224 200FastDigest Eam1104I CTCTTC(1/4)↓ 37 5 5 5 5 3 80°C, 5 min 6 no effect FD0234 50FastDigest Eam1105I GACNNN↓NNGTC 37 5 5 5 5 3 65°C, 5 min 1 no effect FD0244 100FastDigest Ecl136II GAG↓CTC 37 5 5 5 5 1 65°C, 5 min 6 FD0254 100FastDigest Eco105I TAC↓GTA 37 5 5 5 5 3 65°C, 5 min 16 FD0404 50FastDigest Eco130I C↓CWWGG 37 5 5 20 20 3 65°C, 5 min 16 no effect FD0414 200FastDigest Eco147I AGG↓CCT 37 5 5 5 5 3 80°C, 10 min 16 FD0424 100FastDigest Eco31I GGTCTC(1/5)↓ 37 5 5 5 5 3 65°C, 5 min 16 FD0294 100FastDigest Eco32I GAT↓ATC 37 5 5 5 5 2 No 16 no effect FD0304 400FastDigest Eco47I G↓GWCC 37 5 5 5 5 1 80°C, 20 min 16 FD0314 200FastDigest Eco47III AGC↓GCT 37 5 5 5 5 4 65°C, 5 min 16 FD0324 20

FastDigest Eco52I C↓GGCCG 37 5 20 20 5 3 65°C, 5 min 16 FD0334 50FastDigest Eco57I CTGAAG(16/14)↓ 37 15 30 15 15 2 65°C, 5 min 16 no effect FD0344 20FastDigest Eco72I CAC↓GTG 37 5 5 5 5 2 80°C, 10 min 1 FD0364 200FastDigest Eco81I CC↓TNAGG 37 5 5 5 5 2 80°C, 10 min 16 no effect FD0374 50FastDigest Eco88I C↓YCGRG 37 5 5 5 5 2 65°C, 5 min 16 FD0384 200FastDigest Eco91I G↓GTNACC 37 5 5 5 5 2 65°C, 10 min 2 no effect FD0394 200FastDigest EcoO109I RG↓GNCCY 37 5 10 15 5 2 65°C, 5 min 16 FD0264 200FastDigest EcoRI G↓AATTC 37 5 5 20 5 2 80°C, 5min 0.5 FD0274/5 800/2500FastDigest EheI GGC↓GCC 37 5 5 5 5 2 65°C, 5 min 6 FD0444 50FastDigest Esp3I CGTCTC(1/5)↓ 37 5 5 5 5 1 65°C, 10 min 6 FD0454 20FastDigest FaqI GGGAC(10/14)↓ 37 15 15 15 15 3 65°C, 5 min 16 FD1814 20FastDigest FokI GGATG(9/13)↓ 37 5 5 5 5 1 65°C, 5 min 1 no effect FD2144 100FastDigest FspAI RTGC↓GCAY 37 5 5 5 5 4 65°C, 5 min 16 FD1664 20FastDigest FspBI C↓TAG 37 5 15 5 20 3 80°C, 5 min 16 no effect FD1764 50FastDigest GsuI CTGGAG(16/14)↓ 30 15 30 15 15 2 65°C, 5 min 16 FD0464 20FastDigest HhaI GCG↓C 37 5 5 5 5 1 No 16 FD1854 200FastDigest Hin1I GR↓CGYC 37 5 5 5 5 1 65°C, 5 min 16 FD0474 100FastDigest Hin1II CATG↓ 37 5 10 5 10 4 80°C, 5 min 16 no effect FD1834 100FastDigest Hin6I G↓CGC 37 5 10 5 5 2 80°C, 10 min 16 FD0484 200FastDigest HincII GTY↓RAC 37 5 5 5 10 1 65°C, 5 min 16 FD0494 100FastDigest HindIII A↓AGCTT 37 5 5 20 10 3 80°C, 10 min 16 no effect FD0504/5 800/2500FastDigest HinfI G↓ANTC 37 5 5 5 5 2 65°C, 20 min 16 FD0804 400FastDigest HpaII C↓CGG 37 5 5 5 5 3 65°C, 5 min 16 FD0514 200FastDigest Hpy8I GTN↓NAC 37 5 5 5 10 2 80°C, 5 min 16 FD1574 20FastDigest HpyF10VI GCNNNNN↓NNGC 37 5 5 5 5 2 80°C, 5 min 16 FD1734 20FastDigest HpyF3I C↓TNAG 37 5 5 5 10 3 65°C, 5 min 16 no effect FD1884 50FastDigest Kfl I GG↓GWCCC 37 5 5 5 10 5 No 16 FD2164 20FastDigest KpnI GGTAC↓C 37 5 5 5 5 3 80°C, 5 min 16 no effect FD0524 300FastDigest Kpn2I T↓CCGGA 37 5 5 5 5 2 80°C, 5 min 16 FD0534 100FastDigest KspAI GTT↓AAC 37 5 5 5 5 3 65°C, 20 min 0.5 FD1034 50FastDigest LguI GCTCTTC(1/4)↓ 37 5 5 5 5 2 65°C, 5 min 16 no effect FD1934 20FastDigest Lsp1109I GCAGC(8/12)↓ 37 5 5 5 10 2 65°C, 5 min 0.5 no effect FD2074 100FastDigest MauBI CG↓CGCGCG 37 no sites 5 10 5 5 65°C, 5 min 16 FD2084 20FastDigest MbiI CCGCTC(-3/-3)↓ 37 5 5 5 5 3 65°C, 5 min 16 FD1274 100FastDigest MboI ↓GATC 37 5 5 5 10 1 65°C, 15 min 16 FD0814 50FastDigest MboII GAAGA(8/7)↓ 37 5 5 5 5 2 65°C, 5 min 16 FD0824 50FastDigest MlsI TGG↓CCA 37 5 5 30 20 3 80°C, 20 min 16 FD1214 50FastDigest MluI A↓CGCGT 37 5 5 5 15 3 80°C, 5 min 16 FD0564 100FastDigest MnlI CCTC(7/6)↓ 37 5 5 5 5 2 65°C, 5 min 16 no effect FD1074 50FastDigest Mph1103I ATGCA↓T 37 5 5 5 10 3 65°C, 15 min 6 no effect FD0734 100FastDigest MreI CG↓CCGGCG 37 no sites 5 5 5 3 80°C, 5 min 16 FD2024 20FastDigest MspI C↓CGG 37 5 5 5 5 3 No 16 no effect FD0544 400FastDigest MssI GTTT↓AAAC 37 5 5 5 5 2 65°C, 10 min 16 no effect FD1344 100FastDigest MunI C↓AATTG 37 5 5 5 5 3 No 16 no effect FD0754 50FastDigest Mva1269I GAATGC(1/-1)↓ 37 5 5 5 5 3 65°C, 5 min 16 no effect FD0964 50FastDigest MvaI CC↓WGG 37 5 5 5 10 4 No 1 no effect FD0554 200FastDigest NcoI C↓CATGG 37 5 10 10 5 3 65°C, 15 min 16 no effect FD0574/5 100/300FastDigest NdeI CA↓TATG 37 5 5 60 30 3 65°C, 5 min 6 no effect FD0584/5 300/1000FastDigest NheI G↓CTAGC 37 5 15 5 5 5 65°C, 5 min 6 FD0974 100FastDigest NmuCI ↓GTSAC 37 5 5 5 10 2 65°C, 5 min 16 FD1514 20FastDigest NotI GC↓GGCCGC 37 no sites 30 5 10 2 80°C, 5 min 16 FD0594/5 50/150FastDigest NsbI TGC↓GCA 37 5 5 5 5 3 65°C, 15 min 16 FD1224 50FastDigest OliI CACNN↓NNGTG 37 5 5 10 5 3 65°C, 5 min 6 FD1634 20FastDigest PacI TTAAT↓TAA 37 no sites 5 5 5 2 65°C, 10 min 16 no effect FD2204 25FastDigest PaeI GCATG↓C 37 5 5 5 5 5 65°C, 5 min 16 no effect FD0604 20FastDigest PagI T↓CATGA 37 5 10 5 10 3 80°C, 5 min 0.5 FD1284 50FastDigest PdiI GCC↓GGC 37 ND 5 5 5 3 65°C, 15 min 16 FD1524 50FastDigest PdmI GAANN↓NNTTC 37 5 5 5 5 2 65°C, 5 min 16 FD1534 100FastDigest PfeI G↓AWTC 37 5 5 10 5 2 65°C, 5 min 16 FD1784 50FastDigest Pfl 23II C↓GTACG 37 15 15 15 15 3 65°C, 5 min 16 FD0854 50FastDigest PfoI T↓CCNGGA 37 5 5 5 5 3 65°C, 5 min 16 FD1754 20FastDigest Ppu21I YAC↓GTR 37 5 5 5 5 1 65°C, 5 min 6 FD1974 50FastDigest Psp1406I AA↓CGTT 37 5 5 5 5 3 65°C, 5 min 16 FD0944 20FastDigest Psp5II RG↓GWCCY 37 5 5 5 20 2 80°C, 5 min 1 FD0764 50FastDigest PspFI CCCAGC(-1/-5)↓ 37 5 5 5 5 4 80°C, 5 min 16 FD2224 20FastDigest PstI CTGCA↓G 37 5 5 30 5 3 No 16 no effect FD0614/5 800/2500FastDigest PsuI R↓GATCY 37 5 5 5 5 2 80°C, 5 min 16 no effect FD1554 150FastDigest PsyI GACN↓NNGTC 37 5 5 5 5 2 80°C, 5 min 4 no effect FD1334 100FastDigest PteI G↓CGCGC 37 5 5 5 5 3 80°C, 5 min 16 FD2134 20FastDigest PvuI CGAT↓CG 37 5 15 5 5 4 80°C, 5 min 16 FD0624 20FastDigest PvuII CAG↓CTG 37 5 5 5 5 1 No 0.5 no effect FD0634 400FastDigest RruI TCG↓CGA 37 5 5 5 5 5 No 4 FD2154 100FastDigest RsaI GT↓AC 37 5 5 5 5 3 No 16 FD1124 100FastDigest RseI CAYNN↓NNRTG 37 5 5 5 5 4 65°C, 20 min 16 no effect FD2004 20FastDigest SacI GAGCT↓C 37 5 15 30 10 1 65°C, 5 min 16 no effect FD1134 200FastDigest SalI G↓TCGAC 37 5 5 60 5 3 65°C, 10 min 16 FD0644 200FastDigest SaqAI T↓TAA 37 5 5 5 5 4 65°C, 5 min 16 no effect FD2174 50FastDigest SatI GC↓NGC 37 5 5 10 5 3 65°C, 5 min 16 FD1644 20FastDigest ScaI AGT↓ACT 37 5 5 5 5 4 65°C, 10 min 16 no effect FD0434 100FastDigest SchI GAGTC(5/5)↓ 37 5 5 30 5 2 80°C, 5 min 1 no effect FD1374 100FastDigest SdaI CCTGCA↓GG 37 5 5 5 5 3 No 1 no effect FD1194 20FastDigest SduI GDGCH↓C 37 5 5 5 5 2 80°C, 15 min 1 no effect FD0654 50FastDigest SfaAI GCGAT↓CGC 37 no sites 5 60 5 5 80°C, 5 min 6 FD2094 100FastDigest Sfi I GGCCNNNN↓NGGCC 50 no sites 15 15 15 5 No 16 FD1824 150FastDigest SgsI GG↓CGCGCC 37 5 5 5 5 2 65°C, 20 min 16 FD1894 100FastDigest SmaI CCC↓GGG 37 5 5 5 5 1 65°C, 5 min 16 FD0664 200FastDigest SmiI ATTT↓AAAT 37 no sites 20 30 30 2 65°C, 15 min 1 no effect FD1244 200FastDigest SsiI CCGC(-3/-1)↓ 37 5 5 5 5 2 65°C, 5 min 4 FD1794 20FastDigest SspI AAT↓ATT 37 5 5 5 30 2 65°C, 5 min 1 no effect FD0774 100FastDigest TaaI ACN↓GT 65 5 5 5 5 3 No 16 FD1364 20FastDigest TaiI ACGT↓ 65 5 5 5 5 2 No 6 FD1144 50FastDigest TaqI T↓CGA 65 5 5 5 5 4 No 16 FD0674 400FastDigest TasI ↓AATT 65 5 5 5 5 3 No 6 no effect FD1354 100FastDigest TatI W↓GTACW 65 15 15 15 20 5 No 1 no effect FD1294 20FastDigest TauI GCSG↓C 55 15 60 15 30 5 No 16 FD1654 20FastDigest Tru1I T↓TAA 65 5 5 5 10 3 No 2 no effect FD0984 50FastDigest TscAI CASTG(2/-7)↓ 65 5 5 5 5 2 No 6 no effect FD2104 100FastDigest Van91I CCANNNN↓NTGG 37 5 5 5 5 3 65°C, 10 min 6 FD0714 100FastDigest VspI AT↓TAAT 37 5 5 5 30 2 65°C, 5 min 16 no effect FD0914 200FastDigest XagI CCTNN↓NNNAGG 37 5 5 5 20 2 65°C, 5 min 16 no effect FD1304 100FastDigest XapI R↓AATTY 37 5 5 5 10 4 80°C, 5 min 6 no effect FD1384 100FastDigest XbaI T↓CTAGA 37 5 5 5 10 2 65°C, 20 min 16 FD0684/5 300/750FastDigest XceI RCATG↓Y 37 5 5 5 5 2 65°C, 5 min 16 no effect FD1474 50FastDigest XhoI C↓TCGAG 37 5 5 5 10 2 80°C, 5 min 16 FD0694/5 400/1200FastDigest XmaJI C↓CTAGG 37 5 10 5 10 2 No 16 no effect FD1564 20FastDigest XmiI GT↓MKAC 37 5 5 60 5 2 65°C, 5 min 16 FD1484 50

* For digestion with FastDigest Esp3I add 1 mM DTT. For digestion with FastDigest Eco57I, FastDigest AjuI and FastDigest BseMII add 0.01 mM SAM. For digestion with FastDigest BpII and Fastdigest FaqI add 0.05 mM SAM. For digestion with FastDigest BveI add 0.5 µM oligonucleotide. No - thermal inactivation is not possible, spin column purifi cation or phenol/chloroform extration is recommended. ND - not determined.

DNA 1 µg 2 µg 3 µg 4 µg 5 µgFastDigest enzyme 1 µL 2 µL 3 µL 4 µL 5 µL10X FastDigest Green Buffer or 10X FastDigest Buffer

2 µL 2 µL 3 µL 4 µL 5 µL

Total volume 20 µL 20 µL 30 µL 40 µL 50 µL

Scaling up DNA digestion reactions

Increase the incubation time by 3-5 min if the total reaction volume exceeds 20 μL. It is recommended to use a water thermostat since air thermostats exhibit a slow heat transfer to the reaction mixture.

3-5 kb

10 bp

Reaction mixture containing Thermo Scientifi c™ FastDigest™ Green Buffer:1: Loaded into gel well, before electrophoresis2: Separated, after electrophoresis

1 2

FastDigest Green Buffer

Reaction mixture containing Thermo Scientifi c™

1: Loaded into gel well, before electrophoresis2: Separated, after electrophoresis

GeneRuler 1kb Plus DNA Ladder, #SM1331/2/3/4O’GeneRuler 1kb Plus DNA Ladder, #SM1343

GeneRuler 1kb DNA Ladder,#SM0311/2/3/4O’GeneRuler 1kb DNA Ladder, #SM1163

1% A

garo

se

0.5µg/lane, 8cm gel1X TAE, 7V/cm, 45min

8000 30.010000 30.0

5000 30.0

2000 25.0

1000 60.0

500 25.0

250 25.0

750 25.0

2500 25.0

3500 30.04000 30.0

6000 70.0

1500 25.0

3000 70.0

bp ng/0.5µg

0.5µg/lane, 8cm gel1X TAE, 7V/cm, 45min

bp ng/0.5µg

7000 20.010000 20.0

4000 20.0

700 25.0500 75.0

300 25.0

75 25.0

400 25.0

1000 25.0

2000 20.0

3000 20.0

5000 75.0

1500 80.0

20000 20.0

200 25.0

1% A

garo

se

1.7%

Aga

rose

0.5µg/lane, 8cm gel1X TBE, 5V/cm, 1h

bp ng/0.5µg

2000 28.01500 28.01200 28.0

3000 28.0

900 27.0

500 80.0400 30.0

200 30.0

100 30.0

300 30.0

600 27.0700 27.0800 27.0

1000 80.0

GeneRuler 100bp Plus DNA Ladder, #SM0321/2/3/4O’GeneRuler 100bp Plus DNA Ladder, #SM1153

50 80 4020 80 4010 124 62

20 µL/lane, 1X TBE, 7 V/cm

bp ng/20 µL ng/10 µL

200 80 40100 80 40

FastRuler Ultra Low Range DNA Ladder,ready-to-use #SM1233

400 80 40200 80 40

50 80 40

20 µL/lane, 1X TBE, 7 V/cm

FastRuler Low Range DNA Ladder,ready-to-use #SM1103

bp ng/20 µL ng/10 µL1500 80 40850 80 40

FastRuler Middle Range DNA Ladder,ready-to-use #SM1113

20 µL/lane, 1X TAE, 7 V/cm

bp ng/20 µL ng/10 µL

5000 80 40

850 80 40

2000 80 40

400 80 40

100 80 40

FastRuler High Range DNA Ladder,ready-to-use #SM1123

bp ng/20 µL ng/10 µL

10000 80 40

2000 80 404000 80 40

1000 80 40500 80 40

20 µL/lane, 1X TAE, 7 V/cm

4% A

garo

se

1% A

garo

se

1% A

garo

se

2% A

garo

se

Thermo Scientifi c DNA Ladders

© 2014 Thermo Fisher Scientifi c Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientifi c Inc. and its subsidiaries.

To fi nd out more, visit thermoscientifi c.com/fastdigest