品名:DIG-High Prime DNA Labeling and Detection Starter Kit I
檢測(cè)方法:Kit for color detection with NBT/BCIP.
Catalog :11745832910
Pack size :1 kit for 12 labeling reactions and detection of 24 blots 10 ng to 3 µg DNA per assay, blots of 100 cm2
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.
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.
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
1.DIG-High Prime, 5x conc., 50 µl
2.DIG-labeled Control DNA, 20 µl, (5 µg/ml), pBR328 (linearized with Bam HI)
3.DNA Dilution Buffer, 3 x 1 ml
4.Anti-digoxigenin-AP Conjugate, 100 µl
5.NBT/BCIP Stock Solution, conc., 6 x 1 ml
6.Blocking Solution, 10x conc., 4 x 100 ml
7.DIG Easy Hyb Granules, 4 x 100 ml