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Promega Corporation

Sample Types Examined with the DNA IQ™ System

DNA was purified successfully from the following sample types at Promega or by external forensic laboratories. Due to the nature of casework samples (i.e., samples may have been exposed to environmental factors for long periods of time, and the amount of biological material may be limiting), DNA yields may vary, and DNA may not be obtained from all samples. Tissue sources including hair, bone and sperm require proteinase K digestion to obtain reliable amounts of DNA (see Table 2).

Many of these protocols were communicated to Promega by external laboratories. Because these protocols were not performed at Promega, they are intended to be recommendations that might require optimization.

This list will be updated as additional information becomes available. Contact Promega Technical Services (genetic@promega.com) for the latest information on available protocols.

Table 1. Sample Types Examined Using the DNA IQ™ System (see #TB296 or #TB297.)

Sample Type Promega External Comments
Fresh blood Yes Yes Works with the following anti-clotting reagents: EDTA, citrate, heparin, ACD.
See #TB297.
Frozen blood Yes Yes Old blood may produce lower yields.
See #TB297.
Blood stains on: See #TB296 and #TB297 to isolate DNA from stains on solid material.
S&S903
paper
Yes Yes
FTA® paper Yes Yes
Cotton Yes Yes
Blue denim Yes Yes
Black denim Yes Yes
Soil Yes
Leather Yes Yes
Buccal swabs* See #TB297 to isolate DNA from buccal swabs.
The expected DNA yield from a single buccal swab is up to 300ng.
Cotton Yes Yes
Rayon Yes
CEP paper Yes
Swab to FTA®
paper
Yes
Foam swab to
paper
Yes
Cigarette butt Yes Yes Cut paper wrapping into small pieces. Add 200μl of Incubation Buffer from the Tissue and Hair Extraction Kit (for use with DNA IQ™). Incubate at 56°C for 30 minutes. Transfer buffer and sample to a DNA IQ™ Spin Basket (Cat.# V1221). Centrifuge at room temperature for 2 minutes at maximum speed. Discard spin basket. Add 400μl of prepared Lysis Buffer from the DNA IQ™ System, and proceed to Step 5 of Section 4.B of the #TB296.
The filter may form a gel if heated.
Toothbrush Yes Preincubation in Lysis Buffer is recommended. See below.
Differential extractions Yes See the #TB296.
Semen stain (vasectomized) Yes Swabs containing skin epithelial cells were incubated with 250μl of prepared Lysis Buffer for 30 minutes at 95°C.
Envelope Yes Soak in 0.5% SDS before adding 2 volumes of prepared Lysis Buffer.
Urine Yes Pellet cells, discard liquid and add 100μl of prepared Lysis Buffer and 7μl of DNA IQ™ Resin to the cell pellet. Alternatively, add 2 volumes of prepared Lysis Buffer and 7μl of DNA IQ™ Resin to liquid urine.
Proceed with Step 4 of Section 4.C of the #TB296.
Chewing gum Yes Mince gum into small pieces. Preincubation in Lysis Buffer is recommended. See below.
Skin epithelial cells Yes Swabs of skin epithelial cells were incubated in 250μl of Lysis Buffer at 95°C for 30 minutes.
Saliva Yes
Vomit Yes Preincubation in Lysis Buffer is recommended. See below.
Toothpick Yes Preincubation in Lysis Buffer is recommended. See below.
Knit cap Yes Preincubation in Lysis Buffer is recommended. See below.
Glove lining Yes Preincubation in Lysis Buffer is recommended. See below.
Cotton with hand soap Yes Preincubation in Lysis Buffer is recommended. See below.
Cotton with motor oil Yes Preincubation in Lysis Buffer is recommended. See below.
Cotton with hand cream Yes Preincubation in Lysis Buffer is recommended. See below.
Cotton with contraceptive foam Yes Preincubation in Lysis Buffer is recommended. See below.
Cotton with dirt Yes Preincubation in Lysis Buffer is recommended. See below.
Canvas Yes Preincubation in Lysis Buffer is recommended. See below.
Carpet Yes Preincubation in Lysis Buffer is recommended. See below.
Black underwear Yes Preincubation in Lysis Buffer is recommended. See below.
Plastic lining of a sanitary bag Yes Preincubation in Lysis Buffer is recommended. See below.
Pantyhose Yes Preincubation in Lysis Buffer is recommended. See below.
Blood-stained car visor Yes Preincubation in Lysis Buffer is recommended. See below.
Bloodstains from tape Yes Preincubation in Lysis Buffer is recommended. See below.

*Swabs also have been used to collect DNA from a variety of samples and surfaces, including drinking glasses, aluminum cans, phone handles, earrings, car door panels, steering wheels, fingernails and jacket cuffs. Protocols for DNA isolation from these swabs would be similar to the buccal swab protocol.

Samples Requiring Proteinase K Digestion

Table 2. Sample Types That Require Proteinase K Treatment Prior to DNA Purification Using the DNA IQ™ System.

DNA from the following sample types has been successfully purified at Promega or by external forensic laboratories. Due to the nature of casework samples, (i.e., samples may have been exposed to environmental factors for long periods of time, and the amount of biological material may be limiting) DNA yields may vary and DNA may not be obtained from all samples.

The following sample types require a Proteinase K digestion prior to the addition of two volumes of prepared Lysis Buffer to one volume of sample when following the Tissue and Hair Extraction Kit (for use with DNA IQ™) protocol. See #TB307. A separate protocol for bone and antler samples is linked below.

Sample Type Promega External Comments
Tissue
Fresh Yes Yes See #TB307.
Formalin fixed Yes Yes See #TB307.
Hair roots Yes Yes See #TB307.
Hair shafts (mitochondrial DNA) Yes See #TB307.
Bone Yes From pulverized bone samples.
See the Bone protocol for the DNA IQ™ System.
Antler Yes From drill shavings.
See the Bone Protocol for the DNA IQ™ System.
Sweat-stained clothing Yes Soak cuttings in freshly prepared Incubation Buffer/Proteinase K Solution for several hours. Add 2 volumes of prepared Lysis Buffer and 7μl of DNA IQ™ Resin to the supernatant. Proceed to Step 6 of Section 4.B of the #TB296.
Sperm Yes Yes A proteinase K digestion for 5–15 minutes is required if the sperm is not from a differential extraction. Add 2 volumes of prepared Lysis Buffer and 7μl of DNA IQ™ Resin. Proceed to Step 4 of Section 2.C of the #TB296.
Fetal Tissue Yes

Pretreatment of Samples by Soaking in Lysis Buffer

DNA has been isolated from many sample types using the DNA IQ™ System by first soaking the sample or a cutting of the sample in prepared Lysis Buffer prior to adding the DNA IQ™ Resin. The following pretreatment steps have been performed by external laboratories and are recommendations that may require optimization.

1. Prepare Lysis Buffer as described in the #TB296.

2. Soak the sample in prepared Lysis Buffer at 56°C for 30 minutes. Different samples may require different volumes of Lysis Buffer or different incubation temperatures. Add at least enough Lysis Buffer to cover the sample.

3. Remove the matrix from the liquid manually with a sterile forceps. Alternatively, transfer the buffer and sample to a DNA IQ™ Spin Basket (Cat.# V1221), and centrifuge at room temperature for 2 minutes at maximum speed. Discard the spin basket.

4. Proceed to Step 5 of Section 4.B of the #TB296.

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