DNA content of biological samples

Type of sample Amount of DNA

Liquid blood 20,000-40,000 ng/mL
stain 250-500 ng/cm2
Liquid semen 150,000-300,000 ng/mL
Postcoital vaginal swab 10-3,000 ng/swab
Hair (with root)
Plucked 1-750 ng/root
Shed 1-10 ng/root
Liquid saliva 1,000-10,000 ng/mL
Oral swab 100-1500 ng/swab
Urine 1-20 ng/mL
Bone 3-10 ng/mg
Tissue 50-500 ng/mg
 Preservation of Sample for DNA Analysis

1. 75% ethanol

2. 2–5 ml1 liquid blood samples (standard as per many standard protocols) is collected in
pre-sterile EDTA vial. Ethylenediaminetetraacetic acid (EDTA) is a polyprotic acid that
chelate Calcium and other ions, thus irreversibly preventing coagulation of blood in the
tube

Among the most often used preservation method of samples collected for DNA analyses is
freezing.
Freezing at −80 °C or in liquid nitrogen (−196 °C)
short term storage −20 to −28 °C is preferred
Liquid: If liquid blood found on any surface then it is recommended to soak up blood in
sterile cotton swab/cloth or gauge. Allow it to air dry before packaging in paper
envelop, information about exhibit and all details regarding the case should be mentioned on
envelope

WET: Liquid blood from crime scene can also be collected either through sterilized syringe or by
pipette using pre-sterile disposable tip. After this the liquid can be dispensed in EDTA vial.

Wet Blood Stains: If wet blood stains found on any clothing, weapon, small objects or any movable
objects which can be moved easily from the crime scene.
Dried Blood Stains: If dried blood stains found on clothing, weapon, small objects or any movable
objects then it is recommended to preserve whole item directly
Lifting and scraping also

Semen, Seminal Stains: sterile cotton swabs. Swabs should be air dried before packaging.

Tissues, Body Parts or Organs: To prevent microbial decay it is also suggested to use antibiotics
(0.1 mg/ml gentamicin solution).

Bone, Skull, Teeth, or Skeletal Remains:

Hair: Wrapped in a dry paper
 DNA Extraction

(1) disruption of cytoplasmic and nuclear membranes;

(2) separation and purification of DNA from other components of the cell
lysate such as lipids, proteins, and other nucleic acids
(3) concentration and purification of DNA
 DNA Extraction Methods

1. Chromatography-Based DNA Extraction Method

2. EtBr-CsCl Gradient Centrifugation Method
3. Alkaline Extraction
4. Silica Matrices
5. Salting-out method
6. Cetyltrimethylammonium Bromide (CTAB) Extraction
7. Phenol-chloroform method
8. SDS-Proteinase K
9. Silica column-based DNA extraction method
10. Chelex-100 Extraction
11. Magnetic beads
12. Cellulose-based paper
13. Filter paper-based DNA extraction method
 In this method we can separate the DNA based on the solubility of DNA molecules in immiscible
solutions, and that is called the liquid-liquid DNA extraction methods (the phenol chloroform
DNA extraction method is one of these methods).

 Although this method is considered one of the best methods, it is less recommended because of
phenol and chloroform harmful. The quantity and quality of DNA obtained by method are very
good.

 This method is known as a phenol-chloroform and isoamyl alcohol or PCI method of DNA
extraction.

 The major chemicals of PCI DNA extraction methods are Phenol, chloroform and lysis buffer
(contains EDTA, Tris, NaCl, MgCl2, SDS, and other salts).

 The organic component of the technique- phenol and chloroform denature the protein portion of
cells and lysis buffer components help in the cell membrane and nuclear lysis.

 The lysis buffer contains salts like EDTA, NaCl, Tris and SDS.

 In principle, the phenol digests proteins, isoamyl alcohol separates nucleic acid and chloroform
reduces the foaming between interphase.
 In the standard salting-out method, proteins K and RNase are added to them after the
lysis of cells. Saturated NaCl was needed for the proteins to precipitate out of the
solution. The cell samples are centrifuged and then,  DNA is separated by washing it with
detergent like ethanol.

The salting-out DNA extraction method is safer than the phenol-chloroform method. It
depends on the usage of salts such as potassium acetate, sodium chloride, and ammonium
acetate that help in DNA extraction.