Platinum Direct PCR Universal Master Mix

Invitrogen Platinum Direct PCR Universal Master Mix is designed to amplify DNA sequences directly from a variety of samples, without requiring DNA purification. It is ideal for amplification of target DNA from crude samples in applications such as mouse genotyping, CRISPR-edited cell analysis, and target DNA detection from limited materials.

Highlights

  • Universal—One master mix for different sample types, one annealing temperature for different primer sets
  • Convenient—Buffer formulated for primer annealing at 60°C, allowing less PCR optimization
  • Fast—No DNA purification; DNA synthesis at 20 sec/kb
  • Efficient— High inhibitor tolerance; GC enhancer included for amplification of GC-rich sequences
  • Flexible—Optional lysis-and-storage protocol for long-term storage and further testing
  • Reduced pipetting—Master mix format with direct gel-loading dye

The master mix is formulated with Invitrogen Platinum II Taq Hot-Start DNA Polymerase, which is engineered for high inhibitor tolerance, fast DNA synthesis, and high sensitivity. In addition to the enzyme, the master mix consists of dNTPs, other reaction components, and a green dye for direct gel loading. The Lysis Buffer, Proteinase K, and the Invitrogen Platinum GC Enhancer are also included as separate vials in the kit.

Ordering information

Video: How to go straight from sample to PCR with direct PCR

Watch what direct PCR is, how it works, and how to get the most out of a direct PCR kit.

Comparison of conventional approach to PCR vs. direct PCR approach
Click image to enlarge

Figure 1. Comparison of conventional approach to PCR vs. direct PCR approach. The direct PCR approach allows you to skip the sample purification step, minimize sample loss, and reduce hands-on time in the PCR workflow.

Biological samples tested with Platinum Direct PCR Universal Master Mix

Human research tissue

Platinum Direct PCR Universal Master Mix successfully amplifies target DNA sequences from various types of human tissues—saliva, buccal swab, blood (preserved in citrate, heparin, or EDTA), nail, hair, and HeLa S3 cells.

Gel image of PCR products of 0.35 kb and 7.5 kb amplified from various human tissues

Figure 2. Direct DNA amplification from various human research tissues. 0.35 kb and 7.5 kb fragments were amplified from various types of human tissues with the Platinum Direct PCR Universal Master Mix. The lysis protocol was followed for the results shown. The molecular weight marker (M) is Invitrogen TrackIt 1 Kb Plus DNA Ladder.

Mouse samples

Platinum Direct PCR Universal Master Mix successfully amplifies target DNA sequences from various types of mouse tissues—ear, tail, kidney, heart, pancreas, liver, spleen, brain, hair, bone marrow, bone, and tooth.

Gel image of PCR products of 0.3 kb and 3.6 kb amplified from various mouse tissues

Figure 3. Direct DNA amplification from various mouse tissues. 0.3 kb and 3.6 kb fragments were amplified from various types of mouse tissues with the Platinum Direct PCR Universal Master Mix. The lysis protocol was followed for the results shown. Soft tissue was cut into 1 mm pieces, hard tissue like bone was crushed in a ceramic grinder to 1–2 mm. The molecular weight marker (M) is TrackIt 1 Kb Plus DNA Ladder.

Plant samples

Platinum Direct PCR Universal Master Mix successfully amplifies target DNA sequences from plant tissues of various species—wheat leaf, tobacco leaf, birch leaf, maple leaf, magnolia leaf, orchid leaf, Arabidopsis leaf, rose leaf, linden leaf, lilac leaf, strawberry leaf, sild strawberry leaf, Thuja leaf, pine spike, spruce spike, rosehips petal, strawberry, corn seed, peanut, wheat seed, tobacco seed, sunflower seed, and wheat root.

Gel image of 0.3 kb PCR product amplified from various plant tissues

Figure 4. Direct DNA amplification from different plant species and tissue types. A 0.3 kb fragment was amplified directly from leaves, seeds, roots, fruits of plant species listed with the Platinum Direct PCR Universal Master Mix. The direct protocol was followed for the results shown by using 1 mm leaf punch or crushed seed. The molecular weight marker (M) is TrackIt 1 Kb Plus DNA Ladder.

Algae

Platinum Direct PCR Universal Master Mix successfully amplifies target DNA sequences from algae with or without cell wall.

Gel image of 0.11 kb PCR product amplified from algae with or without cell wall

Figure 5. Direct DNA amplification from algae. A 0.11 kb fragment was amplified directly from the single-cell green algae Chlamydomonas reinhardtii with and without the cell wall using the Platinum Direct PCR Universal Master Mix. The lysis protocol was followed for the results shown. The molecular weight marker (M) is TrackIt 1 Kb Plus DNA Ladder.

Bird tissue

Platinum Direct PCR Universal Master Mix successfully amplifies target DNA sequences from various bird tissues—chicken cartilage, chicken meat, and quail feather.

Gel image of 0.24 kb PCR product amplified from various bird tissue

Figure 6. Direct DNA amplification from bird tissue. A 0.24 kb fragment was amplified directly from chicken (cartilage, met) and quail (feather) with the Platinum Direct PCR Universal Master Mix. The lysis protocol was followed for the results shown. The molecular weight marker (M) is TrackIt 1 Kb Plus DNA Ladder.

Bacterial species

Platinum Direct PCR Universal Master Mix successfully amplifies target DNA sequences from gram-positive and gram-negative bacterial species.

Gel image of 0.62 kb and 0.59 kb PCR products amplified from Escherichia coli and Bacillus subtilis

Figure 7. Direct DNA amplification from bacteria. 0.62 kb and 0.59 kb fragments were amplified directly from the gram-negative (Escherichia coli) and gram-positive (Bacillus subtilis) bacteria respectively with the Platinum Direct PCR Universal Master Mix. The lysis protocol was followed for the results shown. The molecular weight marker (M) is TrackIt 1 Kb Plus DNA Ladder.

Zebrafish tissue

Platinum Direct PCR Universal Master Mix successfully amplifies target DNA sequences from various zebrafish tissues—fin, scale, muscle, and brain.

Gel image of PCR product amplified from zebrafish fin
Figure 8. Direct DNA amplification from zebrafish. 0.35 kb and 1 kb fragments were amplified directly from zebrafish tissues with the Platinum Direct PCR Universal Master Mix. Results from the lysis protocol are shown. Successful amplification was observed with fin, scale, muscle, and brain of the zebrafish. The molecular weight marker (M) is TrackIt 1 Kb Plus DNA Ladder.

Drosophila tissue

Platinum Direct PCR Universal Master Mix successfully amplifies target DNA sequences from various Drosophila tissues—whole body, half body, head, and half larva.

Gel image of PCR product amplified from drosophila tissue

Figure 9. Direct DNA amplification from Drosophila. A 0.66 kb fragment was amplified directly from various Drosophila tissues with the Platinum Direct PCR Universal Master Mix. Results from the lysis protocol are shown. Successful amplification was observed with body, head, and larva of Drosophila. The molecular weight marker (M) is TrackIt 1 Kb Plus DNA Ladder.

Blood

Platinum Direct PCR Universal Master Mix successfully amplifies target DNA sequences from human blood preserved in citrate, heparin, or EDTA.

Gel image of 0.35 kb and 7.5kb PCR products amplified from human blood preserved in citrate, heparin, and EDTA

Figure 10. Direct DNA amplification from human blood research samples. 0.35 kb and 7.5 kb fragments were amplified from human blood preserved in citrate, heparin, or EDTA with Platinum Direct PCR Universal Master Mix. The lysis protocol was followed for the results shown. The molecular weight marker (M) is TrackIt 1 Kb Plus DNA Ladder.

FFPE

Platinum Direct PCR Universal Master Mix successfully amplifies target DNA sequences from a mouse kidney FFPE sample.

Gel image of 0.1 kb, 0.2 kb, 0.3 kb, 0.5 kb, 1 kb PCR products amplified from mouse kidney FFPE block
Figure 11. Direct DNA amplification from FFPE samples. Fragments of different lengths (0.1 kb, 0.2 kb, 0.3 kb, 0.5 kb, 1 kb) were amplified from mouse kidney FFPE blocks with the Platinum Direct PCR Universal Master Mix according to the manual recommendations. The molecular weight marker (M) is TrackIt 1 Kb Plus DNA Ladder. Note that FFPE DNA is often fragmented, and sample quality may not allow amplification longer than 0.3 kb.

Human research tissue

Platinum Direct PCR Universal Master Mix successfully amplifies target DNA sequences from various types of human tissues—saliva, buccal swab, blood (preserved in citrate, heparin, or EDTA), nail, hair, and HeLa S3 cells.

Gel image of PCR products of 0.35 kb and 7.5 kb amplified from various human tissues

Figure 2. Direct DNA amplification from various human research tissues. 0.35 kb and 7.5 kb fragments were amplified from various types of human tissues with the Platinum Direct PCR Universal Master Mix. The lysis protocol was followed for the results shown. The molecular weight marker (M) is Invitrogen TrackIt 1 Kb Plus DNA Ladder.

Mouse samples

Platinum Direct PCR Universal Master Mix successfully amplifies target DNA sequences from various types of mouse tissues—ear, tail, kidney, heart, pancreas, liver, spleen, brain, hair, bone marrow, bone, and tooth.

Gel image of PCR products of 0.3 kb and 3.6 kb amplified from various mouse tissues

Figure 3. Direct DNA amplification from various mouse tissues. 0.3 kb and 3.6 kb fragments were amplified from various types of mouse tissues with the Platinum Direct PCR Universal Master Mix. The lysis protocol was followed for the results shown. Soft tissue was cut into 1 mm pieces, hard tissue like bone was crushed in a ceramic grinder to 1–2 mm. The molecular weight marker (M) is TrackIt 1 Kb Plus DNA Ladder.

Plant samples

Platinum Direct PCR Universal Master Mix successfully amplifies target DNA sequences from plant tissues of various species—wheat leaf, tobacco leaf, birch leaf, maple leaf, magnolia leaf, orchid leaf, Arabidopsis leaf, rose leaf, linden leaf, lilac leaf, strawberry leaf, sild strawberry leaf, Thuja leaf, pine spike, spruce spike, rosehips petal, strawberry, corn seed, peanut, wheat seed, tobacco seed, sunflower seed, and wheat root.

Gel image of 0.3 kb PCR product amplified from various plant tissues

Figure 4. Direct DNA amplification from different plant species and tissue types. A 0.3 kb fragment was amplified directly from leaves, seeds, roots, fruits of plant species listed with the Platinum Direct PCR Universal Master Mix. The direct protocol was followed for the results shown by using 1 mm leaf punch or crushed seed. The molecular weight marker (M) is TrackIt 1 Kb Plus DNA Ladder.

Algae

Platinum Direct PCR Universal Master Mix successfully amplifies target DNA sequences from algae with or without cell wall.

Gel image of 0.11 kb PCR product amplified from algae with or without cell wall

Figure 5. Direct DNA amplification from algae. A 0.11 kb fragment was amplified directly from the single-cell green algae Chlamydomonas reinhardtii with and without the cell wall using the Platinum Direct PCR Universal Master Mix. The lysis protocol was followed for the results shown. The molecular weight marker (M) is TrackIt 1 Kb Plus DNA Ladder.

Bird tissue

Platinum Direct PCR Universal Master Mix successfully amplifies target DNA sequences from various bird tissues—chicken cartilage, chicken meat, and quail feather.

Gel image of 0.24 kb PCR product amplified from various bird tissue

Figure 6. Direct DNA amplification from bird tissue. A 0.24 kb fragment was amplified directly from chicken (cartilage, met) and quail (feather) with the Platinum Direct PCR Universal Master Mix. The lysis protocol was followed for the results shown. The molecular weight marker (M) is TrackIt 1 Kb Plus DNA Ladder.

Bacterial species

Platinum Direct PCR Universal Master Mix successfully amplifies target DNA sequences from gram-positive and gram-negative bacterial species.

Gel image of 0.62 kb and 0.59 kb PCR products amplified from Escherichia coli and Bacillus subtilis

Figure 7. Direct DNA amplification from bacteria. 0.62 kb and 0.59 kb fragments were amplified directly from the gram-negative (Escherichia coli) and gram-positive (Bacillus subtilis) bacteria respectively with the Platinum Direct PCR Universal Master Mix. The lysis protocol was followed for the results shown. The molecular weight marker (M) is TrackIt 1 Kb Plus DNA Ladder.

Zebrafish tissue

Platinum Direct PCR Universal Master Mix successfully amplifies target DNA sequences from various zebrafish tissues—fin, scale, muscle, and brain.

Gel image of PCR product amplified from zebrafish fin
Figure 8. Direct DNA amplification from zebrafish. 0.35 kb and 1 kb fragments were amplified directly from zebrafish tissues with the Platinum Direct PCR Universal Master Mix. Results from the lysis protocol are shown. Successful amplification was observed with fin, scale, muscle, and brain of the zebrafish. The molecular weight marker (M) is TrackIt 1 Kb Plus DNA Ladder.

Drosophila tissue

Platinum Direct PCR Universal Master Mix successfully amplifies target DNA sequences from various Drosophila tissues—whole body, half body, head, and half larva.

Gel image of PCR product amplified from drosophila tissue

Figure 9. Direct DNA amplification from Drosophila. A 0.66 kb fragment was amplified directly from various Drosophila tissues with the Platinum Direct PCR Universal Master Mix. Results from the lysis protocol are shown. Successful amplification was observed with body, head, and larva of Drosophila. The molecular weight marker (M) is TrackIt 1 Kb Plus DNA Ladder.

Blood

Platinum Direct PCR Universal Master Mix successfully amplifies target DNA sequences from human blood preserved in citrate, heparin, or EDTA.

Gel image of 0.35 kb and 7.5kb PCR products amplified from human blood preserved in citrate, heparin, and EDTA

Figure 10. Direct DNA amplification from human blood research samples. 0.35 kb and 7.5 kb fragments were amplified from human blood preserved in citrate, heparin, or EDTA with Platinum Direct PCR Universal Master Mix. The lysis protocol was followed for the results shown. The molecular weight marker (M) is TrackIt 1 Kb Plus DNA Ladder.

FFPE

Platinum Direct PCR Universal Master Mix successfully amplifies target DNA sequences from a mouse kidney FFPE sample.

Gel image of 0.1 kb, 0.2 kb, 0.3 kb, 0.5 kb, 1 kb PCR products amplified from mouse kidney FFPE block
Figure 11. Direct DNA amplification from FFPE samples. Fragments of different lengths (0.1 kb, 0.2 kb, 0.3 kb, 0.5 kb, 1 kb) were amplified from mouse kidney FFPE blocks with the Platinum Direct PCR Universal Master Mix according to the manual recommendations. The molecular weight marker (M) is TrackIt 1 Kb Plus DNA Ladder. Note that FFPE DNA is often fragmented, and sample quality may not allow amplification longer than 0.3 kb.

Additional key benefits of Platinum Direct PCR Universal Master Mix

The unique formulation of the Platinum direct PCR master mix's buffer helps reduce tedious optimization step in PCR. The innovative buffer formulation enables annealing of primers at 60°C (universal annealing temperature) regardless of their sequences that follow general primer design rules (Figure 12)​. The buffer also allows successful amplification when calculated Tms are used in the annealing step (data not shown).

Gel image of PCR products amplified from plant, mouse and human tissue using a universal annealing temperature at 60°C

Figure 12. Platinum Direct PCR Universal Master Mix produces PCR products with high specificity and yield when DNA templates of various samples were amplified using the universal annealing temperature at 60°C. Primer sets of varying annealing temperatures (indicated above) were used to amplify five targets from different tissues of plant, mouse, and human. Results of both direct and lysis protocols are shown. The molecular weight marker (M) is TrackIt 1 Kb Plus DNA Ladder.

See the benefits of a universal annealing temperature for PCR

Learn the importance of the annealing step in PCR, how to circumvent optimization steps using a specially formulated PCR buffer, and the benefits of a universal annealing temperature enabled by the buffer.

Due to its innovative buffer, Platinum Direct PCR Universal Master Mix allows for a universal annealing temperature and flexible extension time for co-cycling of all assays.

96 well plate with wells of three different colors to illustrate amplification of three PCR targets at the same time by cocyling
Figure 13. Time saving and assay co-cycling enabled by universal PCR protocol. With the Platinum Direct PCR Universal Master Mix, different PCR assays can be cycled together (co-cycling) using one protocol with a universal primer annealing temperature and the extension time selected for the longest fragment to be amplified (Figure 14).
Gel image of 0.2 kb, 0.6 kb, 1 kb, 1.5 kb, 2 kb PCR products amplified from mouse, plant, and human tissue using a universal PCR protocol with the same extension time

Figure 14. Platinum Direct PCR Universal Master Mix enables cycling of shorter and longer amplicons together. 0.2 kb, 0.6 kb, 1 kb, 1.5 kb, and 2 kb fragments were amplified from samples of different species and tissue types, using the same protocol for all five targets: 94°C denaturation for 15 sec, 60°C annealing for 15 sec, 68°C extension for 40 sec. The extension time was based on length of the longest target.

Platinum Direct PCR Master Mix provides two protocols to amplify target DNA from crude samples. In the direct protocol, a sample of recommended size is added directly to the master mix. In the lysis protocol, a sample of recommended size is lysed first, then a fraction of its supernatant is transferred to the master mix. The direct protocol offers a shorter workflow whereas the lysis protocol allows a flexible workflow with a sample-storage option (Figure 15). Up to 2 kb can be amplified using the direct PCR protocol, and up to 8 kb can be amplified following the lysis protocol (Figure 19).

Illustration of steps in direct vs. lysis protocols with tubes, icons, and thermal cyclers
Figure 15. Direct protocol vs. lysis protocol.

Sample size and amount are critical for success in direct PCR. Low sample amount can cause PCR failure due to insufficient input. On the other hand, large sample size introduces cellular components and debris that can inhibit PCR. Recommended sample sizes are provided in the manual of the Platinum Direct PCR Universal Master Mix.

Recommended sample sizes as cricles illustrated in scale for direct and lysis protocols for direct PCR

Figure 16. Sample size recommendation for the direct and lysis protocol (not rendered for actual size). In general, 0.5–1.0 mm of solid samples works well with the direct protocol. 0.5–2.0 mm is recommended for the lysis protocol although up to 10 mm of sample can work with the lysis protocol. Please refer to the product manual for detailed recommendations on different sample types.

For your convenience, print a sample size guide letter size (8.5” x 11”) or A4 size (210 mm x 297 mm) for solid samples when working with the Platinum Direct PCR Universal Master Mix.

Platinum Direct PCR Universal Master Mix is provided as a green master mix for direct gel loading of PCR products, eliminating tedious steps of dye addition to PCR samples and helping reduce pipetting errors and hands-on time.

Green dye in PCR master mix that separates to yellow and blue dyes in electrophoresis, illustrating direct gel-loading master mix benefits

Figure 17. The green master mix for loading PCR products directly to a gel for analysis. The Platinum Direct PCR Universal Master Mix contains a density reagent and two tracking dyes. DNA migration is easily tracked with two dyes (blue and yellow) that are readily visible during electrophoresis (the lanes for 5 and 15 min in the figure to the right).

Efficient PCR with Platinum Direct PCR Universal Master Mix

PCR run time

Platinum Direct PCR Universal Master Mix is designed with Platinum II Taq Hot-Start DNA Polymerase, which is an engineered enzyme with fast DNA synthesis. Therefore, PCR results can be obtained sooner with the Platinum direct PCR master mix than other direct PCR kits (Figure 17).

Bar graph showing cycling time and ramping time to compare PCR run times of different direct PCR kits

Figure 18. Fast cycling reduces PCR run time. A 1 kb fragment was amplified for 35 cycles using the Platinum Direct PCR Universal Master Mix and direct PCR kits from other suppliers. Cycling times for each kit are shown in purple, while ramping times on the Applied Biosystems ProFlex PCR System (6°C/sec peak block ramp rate) are shown in red.

Amplification length

Platinum Direct PCR Universal Master Mix enables amplification of long sequences directly from the samples.

Gel images of 0.2 kb, 0.35 kb, 0.8 kb, 2 kb, 3.3 kb, 4.5 kb, 7.5 kb PCR products from direct and lysis protocols to show different amplification ranges from the two protocols
Figure 19. Amplification ranges of the direct and lysis protocols using Platinum Direct PCR Universal Master Mix. Fragments up to 2 kb can be amplified with the direct protocol while fragments up to 8 kb can be amplified with the lysis protocol. Results shown are PCR with human blood preserved in heparin. The molecular weight marker (M) is TrackIt 1 Kb Plus DNA Ladder.

High specificity

Platinum Direct PCR Universal Master Mix is designed with Platinum II Taq Hot-Start DNA Polymerase, which incorporates Invitrogen Platinum hot-start technology. This antibody-based hot-start technology offers superior specificity in direct DNA amplification using crude samples.

Gel image of 0.3 kb, 0.5 kb, 1.1 kb, 2.2 kb, 2.9 kb, 3.6 kb and 5.5 kb PCR products to show PCR specificity
Figure 20. High specificity of direct PCR. 0.3 kb, 0.5 kb, 1.1 kb, 2.2 kb, 2.9 kb, 3.6 kb and 5.5 kb targets were amplified from mouse ear following the lysis protocol of the Platinum Direct PCR Universal Master Mix. The molecular weight marker (M) is TrackIt 1 Kb Plus DNA Ladder.

High sensitivity

Platinum Direct PCR Universal Master Mix is designed with Platinum II Taq Hot-Start DNA Polymerase, which is an engineered enzyme with improved sensitivity. Therefore, target sequences can be detected from low sample input using the Platinum direct PCR master mix.

Gel image of different cell numbers (10,000 to 1 cell) to show PCR sensitivity

Figure 21. High sensitivity of PCR from low-sample input. 0.4 kb and 2.2 kb targets were amplified from 5–500 of HeLa S3 cells following the lysis protocol of the Platinum Direct PCR Universal Master Mix. The molecular weight marker (M) is TrackIt 1 Kb Plus DNA Ladder.

AT-rich and GC-rich PCR

Platinum Direct PCR Universal Master Mix allows for amplification of versatile range of targets, from AT-rich to GC-rich. A separate vial of Platinum GC Enhancer is provided for specific and enhanced amplification of targets with high-GC content.

Gel image of PCR products of different GC% (from 29% to 77%) to show AT-rich and GC-rich amplification

Figure 22. Robust direct amplification of AT-rich and GC-rich targets. Fourteen targets of varying GC content were amplified from human buccal swabs following the lysis protocol of the Platinum Direct PCR Universal Master Mix. The Platinum GC Enhancer (included with the kit) was used for targets over 65% GC content. The molecular weight marker is TrackIt 1 Kb Plus DNA Ladder.

Multiplex PCR

Platinum Direct PCR Universal Master Mix allows amplification of multiple targets in a single reaction.

Gel images of one to five PCR products amplified in a single tube using mouse ear, tail, and hair samples
Figure 23. Efficient multiplexing in direct PCR of up to five targets. Fragments of different lengths (0.1 kb to 1.1 kb) were amplified from mouse ear, tail and hair in single-plex to 5-plex reactions. Results from the lysis protocol of the Platinum Direct PCR Universal Master Mix are shown. The molecular weight marker is the Invitrogen TrackIt 100 bp DNA Ladder.

Benchtop stability

Extended stability of the Platinum Direct PCR Universal Master Mix at room temperature enables high-throughput applications. The Platinum hot-start technology of the enzyme allows benchtop stability and high specificity of the kit.

Gel image of PCR products after setting up at room temperature, on ice, or keeping at room temperature for 8 hours to show benchtop stability

Figure 24. Assembled reactions with Platinum Direct PCR Universal Master Mix are stable at room temperature. A 2.2 kb fragment was amplified from mouse tail, following the lysis protocol of the Platinum Direct PCR Master Mix. Reactions were set up at room temperature (25°C) or on ice (4°C), then immediately run for PCR or were left at room temperature (25°C) for 8 hours before PCR. Even after 8 hours at room temperature, the Platinum Direct PCR Master Mix produces results with high specificity and yield. The molecular weight marker (M) is TrackIt 1 Kb Plus DNA Ladder.

Data comparison with other commercial kits using various sample types

Tobacco leaf samples were used with commercially available direct PCR kits to compare yield and specificity of target amplification.

Gel image of 0.15 kb, 0.6 kb, 0.75 kb, 1.6 kb,  and 2.2 kb PCR products amplified from tobacco leaves using Platinum Direct PCR Universal Master Mix and competitor kits

Figure 25. Robust direct DNA amplification from tobacco leaves. Platinum Direct PCR Universal Master Mix (far left panel) provides high specificity and yield across range of DNA fragments (of 0.15 kb, 0.6 kb, 0.75 kb, 1.6 kb, and 2.2 kb) from tobacco leaves, following the direct protocol. The same targets were also amplified using competitor direct PCR kits: (A) Bioline MyTaq Plant PCR Kit, (B) TaKaRa Terra PCR Direct Polymerase Mix, and (C) Sigma-Aldrich REDExtract-N-Amp Plant PCR Kit (lysis protocol). The molecular weight marker (M) is TrackIt 1 Kb Plus DNA Ladder.

Tobacco seed samples were used with commercially available direct PCR kits to compare yield and specificity of target amplification.

Gel image of 0.15 kb, 0.6 kb, 1.6 kb, and 2.2 kb PCR products amplified from tobacco seeds using Platinum Direct PCR Universal Master Mix and competitor kits

Figure 26. Robust direct DNA amplification from tobacco seeds. Platinum Direct PCR Universal Master Mix (far left panel) provides high specificity and yield across range of DNA fragments (0.15 kb, 0.6 kb, 1.6 kb, and 2.2 kb) from tobacco seeds, following the lysis protocol. The same targets were also amplified using competitor direct PCR kits: (A) Sigma-Aldrich REDExtract-N-Amp Plant PCR Kit, (B) Bioline MyTaq Plant PCR Kit, and (C) Roche KAPA3G Plant PCR Kit. The molecular weight marker (M) is TrackIt 1 Kb Plus DNA Ladder.

Mouse ear samples were used with commercially available direct PCR kits to compare yield and specificity of target amplification.

Figure 27. Robust direct DNA amplification from mouse ear. Platinum Direct PCR Universal Master Mix (far left panel) provides high specificity and yield across range of DNA fragments (0.3 kb, 0.5 kb, 1.1 kb, 2.2 kb, 2.9 kb, 3.6 kb, and 5.5 kb) from mouse ear punches, following the lysis protocol. The same targets were also amplified using competitor direct PCR kits: (A) Sigma REDExtract-N-Amp Tissue PCR Kit, (B) Bioline MyTaq Extract PCR Kit, (C) TaKaRa Terra PCR Direct Polymerase Mix, and (D) Roche KAPA Mouse Genotyping Kit. The molecular weight marker (M) is TrackIt 1 Kb Plus DNA Ladder.

Human saliva samples were used with commercially available direct PCR kits to compare yield and specificity of target amplification.

Gel image of 0.2 kb, 0.8 kb, 2 kb, 4.5 kb, and 7.5 kb PCR products amplified from human saliva samples using Platinum Direct PCR Universal Master Mix and competitor kits

Figure 28. Robust direct DNA amplification from human saliva. Platinum Direct PCR Universal Master Mix (far left panel) provides high specificity and yield across a range of DNA fragments (0.2 kb, 0.8 kb, 2 kb, 4.5 kb, and 7.5 kb) from human saliva samples, following the lysis protocol. The same targets were also amplified using competitor direct PCR kits: (A) Sigma-Aldrich REDExtract-N-Amp Tissue PCR Kit, (B) Bioline MyTaq Extract PCR Kit, and (C) TaKaRa Terra PCR Direct Polymerase Mix. The molecular weight marker (M) is TrackIt 1 Kb Plus DNA Ladder.

Human blood research samples were used with commercially available direct PCR kits to compare yield and specificity of target amplification.

Gel images of 0.2 kb, 0.35 kb, 0.8 kb, 2 kb, 3.3 kb, 4.5 kb, and 7.5 kb PCR products amplified from human blood samples using Platinum Direct PCR Universal Master Mix and competitor kits

Figure 29. Robust direct DNA amplification from human blood. Platinum Direct PCR Universal Master Mix (left panels) provides high specificity and yield across a range of DNA fragments (0.2 kb, 0.35 kb, 0.8 kb, 2 kb, 3.3 kb, 4.5 kb, and 7.5 kb) from human blood research samples, following the direct or lysis protocol. The same targets were also amplified using competitor direct PCR kits: (A) Bioline MyTaq Blood PCR kit (direct protocol) and (B) TaKaRa Terra PCR Direct Polymerase Mix (lysis protocol). The molecular weight marker (M) is TrackIt 1 Kb Plus DNA Ladder.

A sample of human cell line was used with commercially available direct PCR kits to compare yield and specificity of target amplification.

Gel images of 0.2 kb, 0.8 kb, 2 kb, 4.5 kb, and 7.5 kb PCR products amplified from human cell cultures using Platinum Direct PCR Universal Master Mix and competitor kits

Figure 30. Robust direct DNA amplification from human cells. Platinum Direct PCR Universal Master Mix (far left panel) provides high specificity and yield across a range of DNA fragments (0.2 kb, 0.8 kb, 2 kb, 4.5 kb, and 7.5 kb) from HeLa S3 cells, following the direct or lysis protocol. The same targets were also amplified using competitor direct PCR kits: (A) TaKaRa Terra PCR Direct Polymerase Mix, (B) Sigma-Aldrich REDExtract-N-Amp Tissue PCR Kit, and (C) Bioline MyTaq Extract PCR Kit. The molecular weight marker (M) is TrackIt 1 Kb Plus DNA Ladder.

Mouse kidney FFPE samples were used with commercially available direct PCR kits to compare yield and specificity of target amplification.

Gel image of 0.1 kb, 0.2 kb, 0.3 kb, 0.5 kb, and 1 kb PCR products amplified from FFPE sample using Platinum Direct PCR Universal Master Mix and competitor kit

Figure 31. Robust in direct DNA amplification from FFPE blocks. Platinum Direct PCR Universal Master Mix (left panel) provides high specificity and yield across a range of DNA fragments (0.1 kb, 0.2 kb, 0.3 kb, 0.5 kb, and 1 kb) from mouse kidney FFPE slides, following the lysis protocol. The same targets were also amplified using TaKaRa Terra PCR Direct FFPE Kit (A). NTC stands for no-template control, and the molecular weight marker (M) is TrackIt 1 Kb Plus DNA Ladder. Note that FFPE DNA is often fragmented, and sample quality may not allow amplification longer than 0.3 kb.

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All trademarks are the property of Thermo Fisher Scientific and its subsidiaries unless otherwise specified. Terra is a trademark of Takara-Clontech Laboratories, Inc. REDExtract-N-Amp is a trademark of Merck KGaA. KAPA and KAPA3G are trademarks of Roche Inc. Bioline MyTaq is a trademark of Bioline Inc.