Platinum Direct PCR Universal Master Mix

Take a direct route to PCR

Invitrogen™ Platinum™ Direct PCR Universal Master Mix is designed to amplify DNA sequences directly from a variety of sample types, without the need for DNA purification. It is ideal for amplification of target DNA from crude samples in applications such as mouse genotyping and analysis of CRISPR-edited cells. The master mix contains a green dye for direct loading of PCR products onto agarose gels.

What makes the master mix universal?One kit that works with different sample types Sample types include human, mouse, plant, fish, bird, bacteria, insect, blood, and formalin-fixed, paraffin-embedded (FFPE) DNA.

One annealing temperature (60°C) for different primer sets

Figure 1. High PCR specificity and yield from direct amplification of various samples using the universal annealing temperature at 60°C following either direct or lysis protocol.

Tobacco leaf

Mouse ear

Mouse tail

Human saliva

Human blood

46°C 52°C 55°C 66°C 68°C

M Dir

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Lysi

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Lysi

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Lysi

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Lysi

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Lysis protocol

Direct protocol

Proteinase K Lysis buffer

DirectPCR

mastermix

Supernatant(1–2 μL)

Sample

Room-temperatureincubation

(≥1 min)

98°Cincubation

(1 min) PCR

+ primers

Long-term storage(4°C to –20°C)

PCR(20 μL)

Sample

PCR

+ primers

DirectPCR

mastermix

PCR

Figure 2. Direct and lysis protocols. Two protocols are available to amplify target DNA directly from crude samples. The direct protocol offers a shorter workflow whereas the lysis protocol allows flexibility and long-term storage.

Flexible workflow

Lysis protocol

Direct protocol

Proteinase K Lysis buffer

DirectPCR

mastermix

Supernatant(1–2 μL)

Sample

Room-temperatureincubation

(≥1 min)

98°Cincubation

(1 min) PCR

+ primers

Long-term storage(4°C to –20°C)

PCR(20 μL)

Sample

PCR

+ primers

DirectPCR

mastermix

PCR

Lysis protocol

Direct protocol

Proteinase K Lysis buffer

DirectPCR

mastermix

Supernatant(1–2 μL)

Sample

Room-temperatureincubation

(≥1 min)

98°Cincubation

(1 min) PCR

+ primers

Long-term storage(4°C to –20°C)

PCR(20 μL)

Sample

PCR

+ primers

DirectPCR

mastermix

PCR

Benefits of Platinum Direct PCR Universal Master Mix include:

• Direct DNA amplification from a variety of sample types

• Less optimization of PCR protocols needed with the universal annealing temperature

• Flexible PCR workflow

• Fast PCR run time (<1 hour)

• Multiplexing with up to 5 targets

• Improved detection with low sample input and GC-rich targets

Primer melting temperatures

For Research Use Only. Not for use in diagnostic procedures. © 2019 Thermo Fisher Scientific Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientific and its subsidiaries unless otherwise specified. COL23636 1019

See more data at thermofisher.com/platinum-direct-pcr

Ordering information

Product Quantity Cat. No.

Platinum Direct PCR Universal Master Mix

100 reactions A44647100

500 reactions A44647500

2,000 reactions A44647200

Long amplicons

Highly sensitive PCR

High benchtop stability

AT-rich and GC-rich PCR

Figure 4. Amplification ranges of the direct and lysis protocols. Up to 2 kb and 7.5 kb fragments can be amplified from human research blood samples with the direct protocol and the lysis protocol, respectively. M = Invitrogen™ TrackIt™ 1 Kb Plus DNA Ladder.

Figure 5. Efficient direct amplification of AT-rich and GC-rich targets. Fourteen targets of varying GC content were amplified from human buccal swabs using the lysis protocol. The Invitrogen™ Platinum™ GC Enhancer is included in the kit for targets over 65% GC. M = TrackIt 1 Kb Plus DNA Ladder.

Direct protocol

M 0.2

kb

0.35

kb

0.8

kb

2 k

b

M

Lysis protocol

M 0.2

kb

0.35

kb

0.8

kb

2 k

b

3.3

kb

4.5

kb

7.5

kb

M

GC content (%)

M 29 30 35 41 57 60 66 67 69 70 71 74 76 77 M

Figure 6. High sensitivity of PCR from low-sample input. A 2.2 kb target was amplified from 5–500 HeLa S3 cells using the lysis protocol. M = TrackIt 1 Kb Plus DNA Ladder.

Figure 7. Stability of reactions at room temperature. A 2.2 kb target was amplified from mouse-tail DNA samples following the lysis protocol. Reactions were set up at 25°C, then immediately run or left for 8 hours at room temperature before PCR. M = TrackIt 1 Kb Plus DNA Ladder.

M 5 cells 50 cells 500 cells M

M 0 hr 8 hr M

2.5

2

1.5

1

.5

0

P A B C D E F G H I J K

PCR

run

time

(hou

rs)

Ramping time

Cycling time

Short PCR run time

Figure 3. Fast cycling reduces PCR run time. A 1 kb fragment was amplified for 35 cycles using the Platinum Direct PCR Universal Master Mix (P) and direct PCR kits from other suppliers (A–K).

P A B C D E F G H JI K