DIG Easy Hyb
英文名称： DIG Easy Hyb
型号:null    产品货号： 11603558001
价格:请致电：010-57128832,18610462672
品牌: roche

DIG Easy Hyb can be used in all membrane hybridization applications using nonradioactive (e.g., digoxigenin-labeled) nucleic-acid probes. DIG Easy Hyb is ready made, and can be used without any additions. It is specially designed for use with DIG kits.

Non-toxic solution, does not contain formamide.

Absence of contaminants: DNase- and RNase-free.

bsp;to establish that the assay is working 
  • Function-tested with the controls provided in the kit (See "Quality" below.) 
  • Sensitive, since the kit can detect as little as 20 fmol of the control oligonucleotide (after it is labeled according to the kit protocol)

    • Product Description

    Brief description
    The kit contains reagents for nonradioactve 3´-end-labeling of oligonucleotides, so they can be used in a "gel mobility shift" assay. It also contains controls, electrophoresis reagents, and an enzyme-labeled antibody to facilitate the detection of DNA-protein complexes.
    Note: The combination of recombinant Terminal Transferase and DIG-11-ddUTP makes the labeling reaction very flexible. It can be used to label the 3-ends of any oligonucleotide (whether it has a 5- overhanging end, a 3-overhanging end or blunt ends). Both single- and double-stranded DNA can be labeled.

    Sample
    Amount: 3.85 pmol or 100 ng 
    Type: Oligonucleotides with 5´-overhanging ends, 3´-overhanging ends, or blunt ends;
    single- or double-stranded DNA fragments (30 - 200 bp)
    Note: Ideally, fragments to be labeled should be between 30 and 100 bp. (See "Notes" below for more details.)

    Time required
    Oligonucleotide annealing and labeling: 10 min
    Formation of oligonucleotide-protein complexes: 25 min
    Electrophoresis: 1 h to overnight, depending on electrophoresis system
    Blotting: 1 - 2 h
    Immunological detection: 2 h
    Exposure to X-ray film or imager: 15 - 40 min 

    Contents

    1. Labeling Buffer (5x conc.), 80 µl
    2. CoCl2 Solution (25 mM), 80 µl
    3. DIG-ddUTP Solution (1 mM Digoxigenin-11-ddUTP), 20 µl
    4. Recombinant Terminal Transferase (400 U/µl), 20 µl
    5. Binding Buffer (5x conc.), 800 µl
    6. Control Oligonucletide (ds 39mer, unlabeled, 0.1 µg/µl, 3.85 pmol/µl), 40 µl
    7. DIG-labeled Control Oligonucleotide (ds 39mer, 0.4 ng/µl, 15.54 fmol/µl), 140 µl
    8. Control Factor Oct2A (25 - 75 ng/µl), 40 µl 
    9. Poly [d(I-C)] (0.1 µg/µl), 200 µl
    10. Poly [d(A-T)] (0.1 µg/µl), 200 µl
    11. Poly L-lysine (0.1 µg/µl in double-distilled water), 200 µl
    12. Loading Buffer without bromophenol blue, 1 ml
    13. Loading Buffer with bromophenol blue, 1 ml
    14. Anti-Digoxigenin-AP (750 U/ml), 40 µl
    15. CSPD (10 mg/ml), 440 µl
    16. Blocking Reagent, 50 g

    Quality

    Function test: Half (50%) of the control oligonucleotide is shifted by the control factor under the conditions described in the protocol. The assay can detect 20 fmol of control oligonucleotide after it is labeled according to the kit protocol.