Production of chromosome region specific DNA sequences and transamination
First Claim
1. In an improved process for transaminating a polynucleotide under aqueous liquid phase conditions, said polynucleotide containing at least one deoxycytidine nucleotide per molecule, in the presence of a bisulfite catalyst and a reactive compound comprising the formula:
- ##STR6## wherein;
X is a divalent radical selected from the class consisting of ##STR7## R is an alkylene radical containing 2 through 12 carbon atoms inclusive, andR1 and R2 are each independently selected from the class consisting of hydrogen and lower alkyl, the improvement which comprises carrying out the transaminating in the presence of a dissolved trihaloacetate chaotrope anion while maintaining a temperature of 20°
to 60°
C. and a pH of 4.5 to 7.5.
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Abstract
Techniques for producing cloned DNA sequences are provided which sequences are complementary to DNA occurring in one selected region of one chromosome of a multi-chromosomal genome, such as the human genome. Such cloned DNA sequences can be labeled and formed into probes by conventional procedures, there are provided methods for making probe compositions which comprise mixed DNA segments derived from such a DNA sequence. An improved DNA sequence transamination procedure is provided utilizing trifluoroacetate chaotrope anions. With high concentrations of low complexity DNA, high levels of transamination are thereby achieved. These segments are covalently bound to fluorophore groups through linking groups that are transaminated preferably chaotropically into the segments.
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Citations
12 Claims
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1. In an improved process for transaminating a polynucleotide under aqueous liquid phase conditions, said polynucleotide containing at least one deoxycytidine nucleotide per molecule, in the presence of a bisulfite catalyst and a reactive compound comprising the formula:
- ##STR6## wherein;
X is a divalent radical selected from the class consisting of ##STR7## R is an alkylene radical containing 2 through 12 carbon atoms inclusive, andR1 and R2 are each independently selected from the class consisting of hydrogen and lower alkyl, the improvement which comprises carrying out the transaminating in the presence of a dissolved trihaloacetate chaotrope anion while maintaining a temperature of 20°
to 60°
C. and a pH of 4.5 to 7.5. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8)
- ##STR6## wherein;
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9. In an improved process for transaminating a polynucleotide under aqueous liquid phase conditions, said polynucleotide containing at least one deoxycytidine nucleotide per molecule, in the presence of a bisulfite catalyst and a reactive compound comprising the formula:
- ##STR8## wherein;
X is a divalent radical selected from the class consisting of ##STR9## R is an alkylene radical containing 2 through 12 carbon atoms inclusive, andR1 and R2 are each independently selected from the class consisting of hydrogen and lower alkyl, the improvement which comprises carrying out the transaminating in the presence of a trifluoroacetate anion while maintaining a temperature in the range of 20°
C. to 60°
C. and a pH in the range of 4.5 to 7.5. - View Dependent Claims (10)
- ##STR8## wherein;
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11. In an improved process for transaminating a polynucleotide under aqueous liquid phase conditions, said polynucleotide containing at least one deoxycytidine nucleotide per molecule, in the presence of a bisulfite catalyst and a reactive compound comprising the formula:
- ##STR10## wherein;
X is a divalent radical selected from the class consisting of ##STR11## R is an alkylene radical containing 2 through 12 carbon atoms inclusive, andR1 and R2 are each independently selected from the class consisting of hydrogen and lower alkyl, the improvement which comprises carrying out the transaminating in the presence of a trichloroacetate anion at a concentration of less than 1M while maintaining a temperature in the range of 20°
C. to 60°
C. and a pH in the range of 4.5 to 7.5. - View Dependent Claims (12)
- ##STR10## wherein;
Specification