比普通DIG随机引物法标记效率高,操作简单,可用于Southern,Northern和斑点杂交以及菌落和噬菌体筛选.一个kit可用于12次标记反应和24次检测反应.组成:5×Dig-GX标记试剂;未标记对照DNA,PBR328;DNA衡释液;抗-Dig-碱性磷酸酶复合物;NBT/BCIP储存液;10x封闭液;Dig定量试纸条;Dig对照试纸条.-20℃保存.
Catalog # Product name / Packsize
11745832910
(1745832) DIG-High Prime DNA Labeling and Detection Starter Kit I
1 kit (12 labeling reactions and 24 detection reactions)
Application
DIG-High Prime labeled probes can be used in Southern-, northern- and dot-blot analysis, as well as colony and plaque hybridizations. This kit offers random-primed labeling of DNA templates with DIG-11-dUTP, alkali-labile, and color detection of the DIG-labeled hybrids. This kit was assembled with convenience in mind, offering a ready-made blocking solution, combined stock solution of NBT/ BCIP, and DIG Easy Hyb granules. The DIG-High Prime mixture includes stabilized Klenow enzyme, nucleotides, primers, and reaction buffer, all in one convenient reagent.
Benefits
The High Prime method of labeling offers many benefits. Plasmid, cosmid, and lambda phage DNA have all been used as templates. We have achieved equal efficiency when labeling 10 ng to 3 μg of template DNA, which may be linear or supercoiled.
Product Description
Sensitivity and specificity: The standard assay uses 1 μg of linearized pBR328. It is usual to expect 0.8 μg labeled DNA after 1 hour of labeling, and 2.3 μg labeled DNA after a 20-hour reaction. In a dot-blot hybridization, 0.1 pg of homologous DNA is detectable after a 16-hour color development (probe concentrations at 20 ng/ml). A single-copy human gene (tPA) is detectable in a Southern blot, loading 1 μg of digested placenta DNA.
Contents
DIG-High Prime, 5x conc., 50 μl
DIG-labeled Control DNA, 20 μl, (5 μg/ml), pBR328 (linearized with Bam HI)
DNA Dilution Buffer, 3 x 1 ml
Anti-digoxigenin-AP Conjugate, 100 μl
NBT/BCIP Stock Solution, conc., 6 x 1 ml
Blocking Solution, 10x conc., 4 x 100 ml
DIG Easy Hyb Granules, 4 x 100 ml
