Kansas State University

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Kansas State University
Department of Plant Pathology

4024 Throckmorton
1712 Claflin Road
Manhattan, KS 66506-5502
785-532-6176
785-532-5692 fax
wgrc@k-state.edu

Wheat Genetics Resource Center
Kansas Wheat Innovation Center

1990 Kimball Ave
Manhattan, KS 66502
785-320-4383

PREHYBRIDIZATION AND 1xbet online sports betting LABELING

Prehybridization

    1. Set hybridization oven to 65°C.
    2. Thaw32P isotope in hood.
    3. Place membrane in hybridization 1xbet online sports betting .
    4. Add 50 mL of prehybridization buffer to each 1xbet online sports betting .
Prehybridization buffer[for 4 probes]
ddH20
125 1xbet online sports betting
20X SSPE
60 1xbet online sports betting
1xbet sports betting Othe
10 1xbet online sports betting
20% SDS
5 1xbet online sports betting

1xbet sports bett[10 mg/1xbet online sports betting ; boil for 5 minutes and set on ice 5 minutes before adding]

1 1xbet online sports betting
TOTAL
200 1xbet online sports betting
  1. Prehybridize membranes for at least 6 hours to overnight.

1xbet online sports betting labeling

    1. Heat water to boiling in a beaker on a hot plate.
    2. Labeling components are
Labeling components
DNA
1.0 µL [20–50 ng]
ddH20
4.0 µL [total volume of H20 and 1xbet online sports betting = 5 µL]
1xbet sports b
1.0 µL
dNTPs [5 mM]
1.5µL

Klenow polymerase

1.0 µL
10X Klenow buffer
1.5µL
dCTP32P
5.0 µL
TOTAL
15.0 µL
  1. Conbine ddH20, oligo primer, and DNA in a 1.5-ml microfuge 1xbet online sports betting , boil for 4 minutes, and place on ice.
  2. Add dNTPs, 10X buffer, Klenow enzyme, and32P. Mix, spin briefly, and allow reaction to go overnight at room temperature or place at 37°C for 2 hours.

Column preparation

  1. Add glass wool to the base of a 1-mL syringe and place in a 15-mL centrifuge 1xbet online sports betting .
  2. Using a transfer pipet, fill the syringe with a solution ofSephadex G-50in TE. Avoid getting air bubbles in the column.
  3. Centrifuge tubes for ~30 seconds, refill tubes with freshSephadex G-50solution, and centrifuge at 1,500 RPM for 4 minutes.
  4. Add 200 µL of TE to each column and centrifuge for 4 minutes at 1,500 RPM.

1xbet online sports betting purification

  1. Add 185 µL of TE to each 1xbet online sports betting .
  2. Cut the cap off the tube and withdraw all of the 1xbet online sports betting solution from the tube with a pipet. Place an empty tube under the syringe and add the 1xbet online sports betting to the top of the column.
  3. Spin the column for 4 minutes at 1,500 RPM.
  4. Discard the column; remove tube with pruified 1xbet online sports betting and recap.
  5. Add a cap-lock to the 1xbet online sports betting , boil for 4 minutes, and place on ice.