New England Biolabs, N3026S, ΦX174 DNA-HaeIII Digest

Related Categories DNA Markers & Ladders Applications DNA Analysis Specification Bases Fragment Mass (ng) bp 1 251 1,353 2 200 1,078 3 162 872 4 112 603 5 58 310 6 102 281/271 7 43 234 8 36 194 9 22 118 10 13 72 Effective Size Range 72bp to 1,353bp Storage Buffer 10 mM Tris-HCl 1 mM EDTA pH 8 @ 25°C FAQ Q: Why is the separation of the lower bands incomplete? A: The low molecular weight ladders contain small fragments and require long run times to resolve the bands effectively. To visualize the smaller bands, a higher percentage gel is helpful (up to 3% agarose, which is most effectively performed using a specially formulated low melt agarose). Smaller bands also tend to diffuse more than larger bands as the electrophoresis is being carried out, so running faster (at higher voltage) can be useful, as long as not too much heat is generated. In most systems, gels of 2% or higher can be run at a voltage of 200 for several hours without excessive heat damage. Q: How can I quantify the amount of DNA in each band of a marker? A: The amount of DNA in each band can be determined by dividing the size (bp) of the band in question by the size (bp) of the uncut DNA molecule and multiplying this number by the total ug of DNA marker loaded on the gel. Q: Can I use Midori Green with the DNA Ladders from NEB? A: NEB DNA Ladders, including Quick-Load Purple 1 kb Plus DNA Ladder (NEB #N0550), are compatible with Midori Green dye with no interference observed. Please follow dye manufacturer's recommendations exactly. Due to low dye intensity, you may need to load between 1 and 2 ug of the DNA ladder to achieve good visualization under standard UV illumination conditions. Q: Can I use SYBR® and/or GelRed® dyes with the DNA Ladders from NEB? A: Because high affinity nucleic acid binding dyes can affect DNA migration during electrophoresis, post-staining of gels with SYBR and GelRed dyes is highly recommended. Post-electrophoresis staining results in superior sensitivity and eliminates the possibility of dye interference with DNA migration. While agarose gels can be precast with these dyes, the migration and/or resolution of some DNA ladders may be affected. Therefore, some DNA ladders may require optimization/diution when run on gels precast with these dyes. For optimum results, please follow the dye manufacturer’s recommendations and protocols carefully, specifically concerning the loading amounts. We recommend using our 1 kb Plus DNA Ladder for Safe Stains (NEB #N0559) when using SYBR or GelRed as precast dyes, as this DNA ladder was specifically optimized for this application. Additionally, avoid using loading dyes that contain SDS, as SDS might cause an abnormal band pattern in precast gels. We recommend using Gel Loading Dye, Purple (6X), no SDS (NEB #B7025) for this application.” Q: Why is my DNA Ladder floating out the wells? Why are there no bands visible in my DNA ladder gel lane? A: If the ready-to-load DNA Ladder or the 6X Loading Dye supplied with the ladder has been frozen at any time, the Ficoll will form a density gradient in the vial upon thawing. Any volume pipetted from the top of the vial will contain only a small amount of density agent and will float out of the wells. To prevent this, mix well upon receipt and before use.