Which DNA Do I Use? How to Choose Your Control and Other DNA Samples

 

DNA double helix molecules and chromosomes.

Today’s Promega Connections blog is written by guest blogger Joliene Lindholm, Promega Technical Services Scientist.

In Promega Technical Services, we are frequently asked questions about choosing among our Human Genomic DNA products. Promega offers DNA that can serve as sources of normal human gene sequences or positive controls where genotype is not critical, and controls for use in genotyping applications like STR analysis. For mouse researchers, we also offer Mouse Genomic DNA.

For Human Genomic DNAs that can serve as sources of normal human gene sequences or as positive controls for experiments where genotype is not critical, we recommend Cat.# G1521, G1471, or G3041. These DNAs are purified from blood pooled from anonymous donors between the ages of 21 and 65. The male and female genomic DNAs have at least six donors per lot, the mixed male and female DNA (Cat.# G3041) has six to twelve donors per lot. This means that there will always be a complex mixture of genotypes that are not present in equal proportions. Because the donors are anonymous, there is no way to know if the same individuals have contributed blood more than once. We expect the genotypes of DNA present in these products will be different in each lot. The human DNA is isolated from pooled blood from healthy individuals and sodium citrate is used to prevent clotting. DNA is isolated the same day the blood is received. The blood is centrifuged to pellet the cells prior to isolating the DNA. The blood is processed using a lysis procedure to isolate white blood cells. The white blood cells are lysed, treated with RNase, and the DNA is purified by phenol extraction, dialysis and isopropanol precipitation. The concentration of the purified DNA is determined by absorbance at 260nm.

  • Each vial contains 100µg of DNA and the lot specific concentration is listed on the tube.
    • Cat.# G1521 is Human Genomic DNA: Female
    • Cat.# G1471 is Human Genomic DNA: Male
    • Cat.# G3041 is Human Genomic DNA: Mixed

We also offer other human genomic DNA products for use as controls in genotyping applications. The 2800M Control DNA is a single-source male human genomic DNA for human STR analysis. Because 2800M is male gDNA, this control DNA is appropriate for use with all PowerPlex® Systems. The genotype is heterozygous at many loci commonly used for forensic analysis. The 2800M Control DNA is available at two concentrations:

  • Cat.# DD7101 contains 25µl at 10ng/µl
  • Cat.# DD7251 contains 500µl at 0.25ng/µl

The K562 DNA High Molecular Weight is purified from a subculture of the human chronic myelogenous leukemia cell line. This DNA is often used as a control in STR analysis; however, some loci display triplex patterns due to culture-induced chromosome rearrangement.

  • Cat.# DD2011 contains 30µg at 0.4–1.0µg/µl. The lot specific concentration is listed on the tube.

The Mouse Genomic DNA is isolated from whole blood of disease-free mice. The strain is Swiss-Webster albino (CFW) mice. Each lot represents a pool of a number of individuals, likely including males and females. The DNA is suitable for Southern blot hybridizations, genomic analysis (including PCR) and genomic library construction.

  • Cat.# G3091 contains 100µg. The lot specific concentration is listed on the tube.

The knowledgeable and experienced scientists in Technical Services are always happy to help you determine the right product that will best suit your needs and applications.

Technical Services Scientist II Joliene earned her Ph.D. in Entomology from the University of Wisconsin-Madison before joining Promega as a Technical Services Scientist. She says “The best thing about my job is learning about all the interesting projects that scientists are investigating; there is always something new and exciting and I get to help them succeed in their goals.” She enjoys dancing and playing ultimate when she is not busy taking care of her menagerie of pets including cockroaches, ducks and rats.

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Promega products are used by life scientists who are asking fundamental questions about biological processes and by scientists who are applying scientific knowledge to diagnose and treat diseases, discover new therapeutics, and use genetics and DNA testing for human identification. Originally, founded in 1978 in Madison, Wisconsin, USA, Promega has branches in 16 countries and more than global distributors serving 100 countries.

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