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Merck

D8661

Sigma-Aldrich

Deoxyribonucleic acid solution from calf thymus

For hybridization

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About This Item

Número de CAS:
MDL number:
UNSPSC Code:
41106310
eCl@ss:
32160414
NACRES:
NA.52

biological source

bovine thymus

Quality Level

grade

for molecular biology

assay

9-11 mg/mL (DNA concentration)

form

solution

shipped in

dry ice

storage temp.

−20°C

General description

A ready-to-use solution of high quality double-stranded template DNA isolated from the thymus of male and female calves. The solution is supplied at a concentration of 9 - 12 mg/ml .

Application

Deoxyribonucleic acid solution from calf thymus is suitable for use as a blocking agent in Southern hybridizations. It was used as negative control in DNA-DNA hybridization experiments using genomic DNA preparations of Vibrio parahaemolyticus and Vibrio alginolyticus. It was used as a standard for DNA quantification by fluorescent assay.
Many factors contribute to the signal-to-noise ratio in nucleic acid hybridizations. These factors include the presence of solvent (formamide), hybridization temperature, length of hybridization, volume of hybridization solution, degree and method of agitation, use of blocking reagents, concentration and specific activity of the probe, use of molecular agents to increase the rate of nucleic acid reassociation, and the degree of stringency used during the washing of the membrane.

In order to decrease any non-specific hybridization of the probe to a substrate, blocking agents must be used. Generally, a combination of blocking reagent, detergent, and denatured, fragmented DNA is used to accomplish this. Sigma offers sonicated, denatured DNA from a variety of species for use as a blocking agent in Northern and Southern blotting and other nucleic acid hybridization techniques.
Many factors contribute to the signal-to-noise ratio in nucleic acid hybridizations. These factors include the presence of solvent (formamide), hybridization temperature, length of hybridization, volume of hybridization solution, degree and method of agitation, use of blocking reagents, concentration and specific activity of the probe, use of molecular agents to increase the rate of nucleic acid reassociation, and the degree of stringency used during the washing of the membrane. 

In order to decrease any non-specific hybridization of the probe to a substrate, blocking agents must be used. Generally, a combination of blocking reagent, detergent, and denatured, fragmented DNA is used to accomplish this. Sigma offers sonicated, denatured DNA from a variety of species for use as a blocking agent in Northern and Southern blotting and other nucleic acid hybridization techniques.

Deoxyribonucleic acid solution from calf thymus has been used in in situ hybridization and microarray hybridization.

Preparation Note

This DNA is phenol-chloroform extracted, ethanol precipitated, and sonicated to produce single-stranded fragments which comigrate with the 587 and 831 base pair marker fragments.

Other Notes

DNA in solution will reanneal on standing at room temperature so it is recommended to boil the solution for 10 minutes and then cool on ice for at least 5 minutes prior to use.

Storage Class

12 - Non Combustible Liquids

wgk_germany

WGK 2

flash_point_f

Not applicable

flash_point_c

Not applicable

ppe

Eyeshields, Gloves, type N95 (US)


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C Booth et al.
Nucleic acids research, 29(21), 4414-4422 (2001-11-03)
We have developed a method that allows quantitative amplification of single-stranded DNA (QAOS) in a sample that is primarily double-stranded DNA (dsDNA). Single-stranded DNA (ssDNA) is first captured by annealing a tagging primer at low temperature. Primer extension follows to
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Theriogenology, 73(4), 523-529 (2010)
Annick Robert-Pillot et al.
FEMS microbiology letters, 215(1), 1-6 (2002-10-24)
We compared the efficiencies of biochemical methods and polymerase chain reaction (PCR) for the identification of Vibrio parahaemolyticus strains. The 122 isolates studied, identified by biochemical tests as V. parahaemolyticus or Vibrio alginolyticus, were tested by R72H PCR assay. The
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