FAQs for λ DNA-BstEII Digest Protocols for λ DNA-BstEII Digest Enzyme Finder Double Digest NEBcutter REBASE

λ DNA-BstEII Digest Catalog # Size Concentration N3014L 750 gel lanes 500 μg/ml N3014S 150 gel lanes 500 μg/ml Download: MSDS PDF Description: The BstEII digest of lambda DNA (cI857 ind 1 Sam 7) yields 14 fragments suitable for use as molecular weight standards for agarose gel electrophoresis (1). The approximate mass of DNA in each of the bands is provided (assuming a 1.0 μg load) for approximating the mass of DNA in comparably intense samples of similar size. Lambda DNA-BstE II Digest visualized by ethidium bromide staining. 1.0% agarose gel. Effective Size Range: 117 bp to 8,454 bp Source: The phage is isolated from the heat-inducible lysogen E. coli λ cI857 S7 and then isolated from the purified phage by phenol extraction and dialyzed.The double-stranded DNA is digested to completion with BstEII, phenol extracted and dialyzed against 10 mM Tris-HCl (pH 8.0) 1 mM EDTA. Reagents Supplied: Gel Loading Dye, Blue (6X) Fragment Size (bp) Mass (ng) 1 8,454 174 2 7,242 149 3 6,369 131 4 5,686 117 5 4,822 99 6 4,324 89 7 3,675 76 8 2,323 48 9 1,929 40 10 1,371 28 11 1,264 26 12 702 14 13 224 5 14 117 2 Concentration: 500 μg/ml Recommended Load: 1 μg Storage Conditions Storage Conditions: 10 mM Tris-HCl 1 mM EDTA pH 8.0 @ 25°C Storage Temperature: -20°C Notes General notes: 1X Gel Lodaing Dye, Blue: 2.5% Ficoll-400 11 mM EDTA 3.3 mM Tris-HCL (pH 8.0@25°C) 0.017% SDS 0.015% bromophenol blue Usage notes: For long term storage, store at -20°C. If samples need to be diluted, use TE or other buffer of minimal ionic strength. DNA may denature if diluted in dH20 and subsequently heated. Temperatures > 60°C may cause denaturation. The cohesive ends of fragments 1 and 4 may be separated by heating to 60°C for 3 minutes. FAQs What are the overhangs on the DNA ladder fragments? Can I end-label them using the T4 polynucleotide kinase (PNK)? What about the Klenow fragment? How can I quantify the amount of DNA in each band of a marker? Why does my Lambda DNA molecular weight marker appear smeared and not represent the correct banding pattern? References Daniels, D.L. et al (1983) In Appendix II: Complete Annotated Lambda Sequence. R.W. Hendrix, J.W. Roberts, F.W. Stahl and R.A. Weisberg (Eds.), Lambda-II, pp. 519-676.  Forster, A.C. et al. (1985) Nucl. Acids Res., 13, 745-761. Reagents Sold Separately Gel Loading Dye, Blue (6X) Companion Products 1 kb DNA Ladder 100 bp DNA Ladder 2-Log DNA Ladder (0.1–10.0 kb) PCR Marker TriDye™ 1 kb DNA Ladder TriDye™ 100 bp DNA Ladder TriDye™ 2-Log DNA Ladder (0.1 - 10.0 kb)