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Many DNA extraction methods can be laborious and time consuming or involve the use of hazardous chemicals. MyTaq™ Extract-PCR Kit offers a rapid, easy and safer alternative for the extraction and amplification of DNA from a variety of tissue types. MyTaq Extract-PCR Kit is particularly suited to solid tissues such as mouse tail or mouse ear. The DNA extractions are performed in a single-tube, without the need for multiple washing steps, greatly reducing the risk of sample loss and contamination.
The extracted DNA is amplified in a proprietary buffer system using MyTaq HS Red Mix, the latest generation of very high-performance polymerase unique to Meridian. To further reduce non-specific amplification, MyTaq HS uses antibody hot-start technology. The advanced formulation of MyTaq HS Red Mix allows fast cycling conditions to be used, greatly reducing the reaction time without compromising PCR specificity or yield.
The rapid MyTaq Extract-PCR Kit maximizes sensitivity while minimizing contamination risks to deliver improved success rates in applications such as mouse DNA characterization. The single tube lysis protocol and MyTaq HS Red Mix maximize sensitivity, while minimizing contamination risks and significantly reduce reaction times as well as delivering improved success rates in protocols such as mouse DNA characterization
MyTaq Extract-PCR was used to extract and amplify genomic DNA from 3 mg pieces of mouse tail. A two-fold serial dilutions of the extract was prepared, then amplified (Lanes 1 – 11) used in PCR reactions containing MyTaq HS Mix and primers for amplification of a 1 kb fragment from the mouse γ-actin (Lanes 1 – 11). Marker is EasyLadder I (M). The results illustrate increased yield across all template input amounts versus all other kits tested.
Comparison of amplification of fragments of the mouse cortexin-1 gene (CTXN1). The MyTaq Extract-PCR Kit and a corresponding kit from Supplier S were used to extract and amplify genomic DNA from 3 mg pieces of mouse tail according to the manufacturers’ instructions. Two-fold serial dilutions (Lanes 1 – 12) were used for the amplification of a 1 kb fragment (A) and a 2 kb fragment (B) from the mouse CTXN1 gene. The extraction protocol and PCR conditions used in each case were those recommended by each supplier. Marker - EasyLadder I (M). The results illustrate increased yield across all template input amounts versus the kit from Supplier S.
Reagent |
100 Reactions |
500 Reactions |
Buffer A |
2 x 1 mL |
10 x 1 mL |
Buffer B |
1 x 1 mL |
5 x 1 mL |
MyTaq HS Red Mix, 2x |
1 x 1.25 mL |
5 x 1.25 mL |
2x
Meridian operates under ISO 13485 Management System. MyTaq Extract-PCR Kit and its components are extensively tested for activity, processivity, efficiency, heat activation, sensitivity, absence of nuclease contamination and absence of nucleic acid contamination prior to release.
All components should be stored at -20°C upon receipt for optimum stability. Repeated freeze/thaw cycles should be avoided.
When stored under the recommended conditions and handled correctly, full activity of the reagents is retained until the expiry date indicated on the outer box label.
Shipped on Blue Ice.
Observation | Recommended Solution(s) |
No or low PCR yield | Enzyme concentration too low – increase the amount of enzyme in 0.5 U increments. |
Primers degraded – check quality and age of the primers. | |
Magnesium concentration too low – increase concentration in 0.25 mM increments with a starting concentration of 1.75 mM. | |
Primer concentration not optimized. Titrate primer concentration (0.3-1 µM); ensuring that both primers have the same concentration. | |
Template concentration too low – Increase concentration of template. | |
Perform a positive control to ensure that the enzyme, dNTPs and buffers are not degraded and/or contaminated. | |
Multiple Bands | Primer annealing temperature too low. Increase annealing temperature. Primer annealing should be at least 5°C below the calculated Tm of primers. |
Prepare master mixes on ice or use a heat-activated polymerase. | |
For problems with low specificity. Try adding 3% DMSO (not supplied) to improve specificity. | |
Smearing or artifacts | Template concentration too high. Prepare serial dilutions of template. |
Too many cycles. Reduce the cycle number by 3-5 to remove non-specific bands. | |
Enzyme concentration too high - decrease the amount of enzyme in 0.5 U increments. | |
Extension time too long. Reduce extension time in 0.5-1 minute increments. |
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