RECIPES: Digoxigenin Labeling Mixes, DNA and RNA

Recipe for making dig-dUTP / dNTP mix for DNA - PCR probe synthesis.. 5/11/96

                           desired     ul of
                         final conc    stock
dATP    100 mM stock        2 mM        3.04
dGTP    100 mM stock        2 mM        3.04
dCTP    100 mM stock        2 mM        3.04
dTTP    100 mM stoc     0.825 mM        1.26
dig dUTP  1 mM stock    0.165 mM        25    (volume of stock supplied)
				  H20  117
                    			152  total volume
dig-dUTP:dTTP  1:5
0.165 mM    0.825 mM
dig-dUTP    dTTP

dig-dUTP    25nmol/25ul = 1 mM          1mM > 0.165 mM
 151.5 ul total volume to dilute dig-11 dUTP to stock to 0.165 mM

Recipe for making dig-UTP / NTP mix for riboprobe (5/16/96)

		       desired     ul of
                     final conc    stock
ATP 100 mM stock        10 mM       7.2
GTP 100 mM stock        10 mM       7.2
CTP 100 mM stock        10 mM       7.2
UTP 100 mM stock        6.5 mM      4.7
dig UTP 10 mM stock     3.5 mM     25.0  (volume of stock supplied)
                              H20  20.7
                                   72.0 total volume

Sources...

DNA:
set of dATP, dGTP, dCTP, dUTP from Boehringer, separate vials, 10
umol / 100 ul each, # 1-277-049 ($72)

individual dNTPs in my lab have been... (I will prob. order from B-M)
dATP  Pharmacia, 100 mM in 100 ul, 27-2050-01
dCTP  Pharmacia, 100 mM in 100 ul, 27-2060-01
dGTP  Pharmacia, 100 mM in 100 ul, 27-2070-01
dTTP  Pharmacia, 100 mM in 100 ul, 27-2080-01

dig-11-dUTP, Boehringer-Mannheim, 25 nmol / 25 ul, # 1-093-088
($133)

RNA:
set of ATP, GTP, CTP, UTP from Boehringer, separate vials, 20 umol /
200 ul each, # 1277-057 ($131)

dig-11 UTP, Boehringer-Mannheim, 250 nmol / 25 ul, # 1209-256
($134)