Are you spending too much time and money extracting and purifying DNA samples for PCR analysis? Now you can avoid the messy cleanup steps and go direct to DNA amplification from a wide range of difficult sample types.
USB® products from Affymetrix®, the leader in enzymatic PCR cleanup, has introduced a new mutant Taq polymerase, VersaTaq Direct PCR Polymerase, which enables direct amplification from samples containing potent PCR inhibitors. Here are key features of this versatile enzyme:
- Eliminates costly extraction and purification steps
- Shorter experiment times leading to faster data output
- Amplify limited amounts of DNA
- Increased sensitivity
- Can be used with a variety of sample sources
Save money and time by avoiding messy cleanup steps
Taq DNA polymerase activity is blocked by PCR inhibitors, like hemoglobin and humic acid, which are found in blood and soil. Thus, a standard Taq DNA polymerase requires procedures to extract and purify DNA prior to PCR. These steps can be time-consuming, may result in the loss of the starting material, and sometimes failed amplification. Unlike standard Taq DNA polymerases, VersaTaq Direct PCR Polymerase is resistant to many potent PCR inhibitors. This mutant Taq jump starts your setup directly to the PCR step, eliminating costly extraction steps. With the VersaTaq Direct PCR Polymerase protocol, you start directly with PCR setup in a few simple steps.
Many sample type options to choose from
VersaTaq Direct PCR Polymerase works with a wide range of samples that would pose an issue for a regular Taq polymerase. Moreover, this enzyme has 5’->3’ exonuclease activity, making it suitable for hydrolysis probe-based detection chemistries. VersaTaq Direct PCR Polymerase can be used in end-point PCR applications directly with many potent PCR inhibitors such as those found in:
- Whole blood
- Blood spots on sample collection cards
- Buccal swabs
- Animal tissue
- Plant tissue
- Crude extracts
Whole blood just got easier to work with
Hemoglobin, a known inhibitor of polymerase activity in PCR reactions, poses a serious challenge for clinical and forensic PCR applications. VersaTaq Direct PCR Polymerase amplifies DNA from as high as 40% human whole blood. It also works with a wide range of targets, including those that are rich in GC content with the use of PCR enhancers such as betaine (Figure 2).
Figure 2. Amplification of targets directly from human whole blood. Targets were amplified from different concentrations of human whole blood in the presence of varying types of anticoagulants. Lanes 1-6: NUMB 455 bp; Lanes 7-12: NUMB 1.8 kb; Lanes 13-15: HRES1 488 bp corresponding band is an 81% GC amplicon. 1 M betaine added to reactions with high GC targets.
A cleaner approach to soil samples
Crude soil samples collected for PCR applications in environmental and forensic fields pose significant challenges. Extensive purification of samples is required due to the presence of humic acid, a byproduct of decomposed organic matter commonly found in soil. Humic acid inhibits the enzymatic activity of standard Taq polymerase. VersaTaq Direct PCR Polymerase easily overcomes the inhibitory effect of humic acid when used directly with crude soil samples, enabling amplification of a variety of targets including those with high GC content (Figure 3).
Figure 3. Amplification of diverse targets in presence of humic acid by VersaTaq Direct PCR Polymerase. Targets were amplified from 5 ng of genomic DNA in the presence of varying concentrations of humic acid. Lanes 1-2: NUMB 455 bp; Lanes 3-4: NUMB 1.8 kb; Lanes 5-6: HRES1 488 bp; Lanes 7-8: SIM2 1.2 kb. 1 M betaine added to reactions with high GC content.
It works hard, so you don’t have to
VersaTaq Direct PCR Polymerase is specially engineered to have increased sensitivity that allows you to amplify limited quantities of DNA in the presence of various PCR inhibitory substances found in difficult samples. For example, VersaTaq Direct PCR Polymerase can amplify DNA from human or animal feces, which normally pose a challenge due to the presence of potent PCR inhibitors such as bile salts (Figure 4). Additionally, this versatile enzyme works directly with different tissue samples, such as those isolated from genetically modified mice, allowing you to skip the DNA extraction step to produce faster results (Figure 5).
Figure 4. Amplification of targets in presence of bile salts by VersaTaq Direct PCR Polymerase. Targets were amplified from 5 ng of genomic DNA in the presence of varying concentrations of bile salts. Lanes 1-2: NUMB 455 bp; Lanes: 3-4 NUMB 967 bp; Lanes: 5-6 HRES1 488 bp. 1 M betaine added to reactions with high GC content.
Figure 5. Amplification of targets directly from mouse tail with VersaTaq Direct PCR Polymerase. Targets were amplified from 1 mm mouse tail clip added directly into PCR reaction. Lane 1: NUMB 483 bp; Lane 2: NUMB 1.8 kb.
Skip DNA extraction and go direct to PCR with VersaTaq Direct PCR Polymerase from Affymetrix, USB.