——More Genotyping, Less Time and Fewer Reagents
PCR-based Genotyping and other experiments often require the purification of genomic DNA from samples. When there are many samples to be tested, the process of purifying genomic DNA is time-consuming, repetitive, and costly. YEASEN Biology has developed a series of Direct PCR Kit products by modifying Taq DNA polymerase and optimizing the PCR reaction system, eliminating the need for genomic purification.
Clever PCR: Direct PCR
YEASEN's Direct PCR Kit starts directly from the sample, using minimal amounts of raw samples (Figure 1-A) or their lysate products (Figure 1-B) without genomic purification as templates for PCR-based Genotyping (Figure 1). Compared to conventional PCR-based Genotyping, the Direct PCR Kit uses less sample, is easy to operate, has a low experimental cost, and allows for high-throughput sample detection.
Fig.1 Direct PCR Amplification Workflow
A.Direct Method: Take a small amount of the sample and directly add it to the PCR Master Mix for PCR identification.Technical Advantages
Direct: No need for time-consuming and expensive DNA purification.
Fast: Template preparation completed in 10 min, and PCR reaction completed in as little as 30 min.
Simple: Samples can be lysed without the need for cutting or grinding, and the PCR Mix is in a pre-mixed form, reducing the number of pipetting steps.
Saving: Small amount of materials used.
High-throughput: Lysis reactions can be completed in a 96-well plate.
Inhibition-resistant: Tolerant to various animal and plant samples.
Direct PCR Kit Experimental Process
This series of products uses a unique protease lysis system to lyse samples. The lysis solution can be directly used as a template and paired with the PCR Master Mix provided in the kit for gene amplification.
Fig.2 Direct PCR experiment process of mouse tissues
Fig.3 Process of direct PCR experiment on plant tissue
Direct PCR Product Selection Table
|
Product Series |
Mouse Direct PCR Second Generation |
Mouse Direct PCR First Generation |
Blood Direct PCR |
Plant Direct PCR |
|
Product Number |
10189ES |
10185ES |
10188ES |
10187ES |
|
Specifications |
20 T/50 T/200 T/500 T |
50 T/200 T |
20 T/50 T/200 T/500 T |
50 T/200 T |
|
Features |
||||
|
Amplicon Length |
≤5 kb |
≤1 kb |
≤8 kb |
≤1 kb |
|
Extension Time (s/kb) |
3-20 s |
30 s |
≤2 kb: 3-5 s ≤8 kb: 10 s |
60 s |
|
Applicable Organisms |
Mice, Rats |
Mice, Rats |
Humans, Mice, Goats, Chickens, Pigs, etc. |
Rice, Corn, Tobacco, Rapeseed, Wheat, Soybeans, etc. |
|
Applicable Tissue/Material Types |
Mouse tail, Mouse ear, Toes (with muscle), and other organs |
Mouse tail, Mouse ear, Toes (with muscle), and other organs |
Fresh blood containing common anticoagulants like EDTA, heparin, citrate, refrigerated (frozen) blood, and commercial dried blood spots like Whatman903® and FTA® |
Tender leaves, Old leaves, Seedlings, Tender stems |
|
Sample Usage |
Tissue: 5 mg Mouse tail: 1-5 mm |
Tissue: 5-10 mg Mouse tail: 1-5 mm |
Whole blood: 0.5%-20% Whatman® dried blood spots: 1 mm2 |
Leaf: 1-10 mm Seed: 1-3 mm |
|
Genomic Release Time |
5-10 min |
15 min |
0-3 min |
0-10 min |
|
Product End Structure |
Blunt ends |
Blunt ends |
Blunt ends |
Blunt ends |
|
Annealing Temperature |
Tm-(2~5)℃ |
Tm-(2~5)℃ |
Tm-(1~2)℃ |
Tm-(2~5)℃ |
|
GC Clipping Range |
29-70% |
30-70% |
30-75% |
40-65% |
|
5'-3' Exonuclease Activity |
Yes |
Yes |
Yes |
Yes |
|
Applications |
||||
|
Genotyping |
Yes |
Yes |
Yes |
Yes |
|
Multiplex PCR |
Up to 5x |
|
Up to 3-4x |
|
|
Direct Sample Amplification |
Yes |
Yes |
Yes |
Yes |
|
Other Features |
||||
|
Electrophoresis Indicator |
Colorless |
Blue |
Colorless |
Blue |
|
Hot Start |
Yes |
Yes |
Yes |
Yes |
|
Premixed/Kit |
Kit (with Mix) |
Kit (with Mix) |
Kit (with Mix+buffer) |
Kit (with Mix) |
Product Advantages
The modified Taq DNA Polymerase is the guarantee of Direct PCR: It has super tolerance to inhibitors.
When non-purified samples are used as templates, many components that inhibit PCR reactions are introduced. It is difficult to achieve ideal results, or even impossible to perform PCR amplification, with ordinary PCR reaction systems and Taq DNA polymerase. The PCR Mix in this series of kits is a formula-optimized premix, using a modified Taq DNA Polymerase, which increases the binding ability to template DNA and has higher tolerance to common PCR inhibitors, thus having a broader template adaptability.
Fig.4 The PCR Mix in the Direct PCR Kit has a higher tolerance to impurities. The same amount of template is used for PCR with different PCR Mixes, and various concentrations of PCR reaction inhibitors are added to the system. The amount of PCR products under different conditions is compared.
YEASEN has developed a series of products for different samples:
- I. Rodent-specific type: Mouse Tissue Direct PCR Kit Plus (10189ES)
Applicable samples: Mouse tissues (such as mouse tail, mouse ear, mouse toe, etc.)
- High amplification efficiency (length, speed, GC compatibility)
Fig.5 The Mouse Tissue Direct PCR Kit Plus amplifies lysates from different tissues. 1: Plasmid; 2: Mouse Ear; 3: Mouse Tail; 4: Liver. Lysis conditions: 70°C lysis for 10 min, 95°C inactivation for 5 min. The target fragments are 1 kb, 2 kb, 3 kb, and 6 kb, with extension times set to 3, 5, 10, and 20 sec/kb, respectively. M: 10510ES60 (YEASEN).
- Broad template compatibility (10189ES)
Fig.6 The Mouse Tissue Direct PCR Kit Plus was used to directly lyse (70°C for 10 min) and amplify human 293 cells (1), saliva (2), and scalp tissue (3). All samples showed clear target bands, M: 10510ES60 (YEASEN).
- II. Multi-species Blood Universal Type: Blood Advanced Direct PCR Kit (10188ES)
- Amplification length up to 8 kb
Fig.7 DNA fragments of varying lengths were amplified directly from human whole blood using the Blood Advanced Direct PCR Kit. The blood samples were stored at -20℃ with EDTA as an anticoagulant. The blood template concentration was 10%, and the extension time was set to 10 sec/kb. M: 10510ES60 (YEASEN).
- High Tolerance
Fig.8 A: The Blood Advanced Direct PCR Kit is used to amplify a 2 kb DNA fragment with a GC content of 71% directly from human EDTA anticoagulated whole blood. The blood template concentration is gradient-set from 0.5-50%, and the extension time is set to 10 sec/kb. B: The Blood Advanced Direct PCR Kit is used to amplify a 2.2 kb DNA fragment with a GC content of 67% directly from mouse EDTA anticoagulated whole blood. The blood template concentration is set from 0.5-50%, and the extension time is 10 sec/kb. M: 10510ES60 (YEASEN).
- III. Plant-specific type: Plant Tissue Direct PCR Kit (10187ES)
- Verified with multiple types of plant samples
Fig.9 Direct amplification results from leaf tissues of different types of plants. M: 100 bp DNA ladder. C: Purified genomic DNA from the corresponding plants as a template. 1-2: Leaf tissue lysate as a template. Number of cycles: 30 cycles.
Customer Case
- I. Mouse Genotype Identification
A biological company in Guangzhou:
Fig.10 Amplification of mouse tail using the Mouse Direct PCR Kit (10189ES). Target gene: 500 bp; Lysis time: 10 min; PCR reaction system: 25 μL, extension time: 5 sec/kb.
- II. Rice Gene Direct Amplification
XX University
Fig.11 The Plant Direct PCR Kit (10187ES) is used to amplify genes related to rice.
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Product Specification |
|
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|
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|
Rapid PCR upgrade, can be colony PCR and suitable for complex template amplification |
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