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.