Skip Lengthy DNA Isolation with Direct PCR!

news February 16 2018

Since its dawn in the 1980s, PCR has become a mainstay in most clinical and biological research laboratories. While Real-Time PCR is invaluable in gene expression analysis and determination of gene copy number, end-point PCR remains the method of choice for many screening applications, such as genotyping, paternity testing, pathogen screening, and transgene detection, to name a few.

Conventional PCR requires the isolation of high quality inhibitor-free DNA from the tissue type or organism under investigation. DNA extraction and purification (e.g., to remove PCR inhibitors) can be tedious, time-consuming, and expensive, and may require the use of toxic chemicals. Furthermore, while the PCR reaction itself is often straightforward, obtaining sufficient template DNA may be challenging, especially when the source material is limited. Direct PCR provides an excellent solution to all of these problems!

So What Is Direct PCR?

During Direct PCR, amplification is performed directly using unpurified sample material as the PCR template e.g., whole blood or plant tissue or crude extracts. By bypassing the need to isolate and purify DNA, the workflow is greatly simplified, as illustrated in Figure 1.

Direct PCR exploits modified polymerases and buffers that facilitate robust DNA amplification despite the presence of PCR inhibitors in the unpurified sample.

Direct PCR will save you time and money, and allows you to circumvent the risk of sample loss when attempting to isolate DNA from limited amounts of starting material.


Figure 1: Direct PCR workflow with Bioline’s My-Taq™ Plant-PCR Kit

Bioline’s Direct PCR Kits

Nordic BioSite supplies Bioline’s Direct PCR kits for the following sample types:

✔ Plant tissue
✔ Whole blood
✔ Mammalian tissue e.g., mouse tail, ear

All of Bioline’s Direct PCR kits incorporate MyTaq HS DNA Polymerase, a next-generation hot-start polymerase that delivers highly specific PCR amplification. MyTaq HS also exhibits increased affinity for template DNA, resulting in high PCR product yields for most challenging templates. Because MyTaq HS DNA Polymerase has been developed to perform more robust amplification than other commonly used polymerases, it performs reliably even in the presence of PCR inhibitors.

MyTaq™ Plant-PCR Kit

This kit is highly suitable for fast, specific and direct PCR from leaf samples from a wide range of plant species, including:

Tomato, rice, sugarcane, maize, potato, soya, wheat, barley, cashew, oak, dwarf umbrella tree, Australian laurel, red-fruit saw-sedge, blueberry lily, she-oaks, coastal rosemary, burrawang, eucalyptus/gum tree, seaweed/crayweed, coastal banksia, bottle brush and honey gem.

Applications

✔ Plant genotyping
✔ Mutation detection
✔ Transgene detection
✔ Knockout analysis

Advantages of MyTaq™ Plant-PCR Kit

✔ Novel buffer system completely circumvents sample processing, DNA extraction and purification
✔ Advanced formulation allows fast cycling conditions, without compromising PCR specificity or yield
✔ Kit is specifically developed to tolerate common PCR inhibitors, including polyphenolics and polysaccharides

In addition to the plant species listed above, customers of the MyTaq™ Plant-PCR Kit have also reported success in wheat, rosemary, cotton, tobacco, citrus, kangkong, Arabidopsis, cranberry, primrose, blueberry, American bellflower and several grass and fungus species. If you don’t see your species listed here, get in touch with Nordic BioSite and we will assist you.

Poster: My-Taq™ Plant-PCR kit for plant genome analysis

MyTaq™ Blood-PCR Kit

This kit is suitable for fast, specific and direct PCR from human and animal whole blood and buccal cell lysates.

Applications

✔ Single nucleotide polymorphism (SNP) genotyping
✔ Genetic testing
✔ Pathogen detection
✔ Blood screening
✔ Paternity testing

Advantages of MyTaq™ Blood-PCR Kit

✔ The unique buffer system and highly developed polymerase overcome the effects of common blood-borne PCR inhibitors e.g., anticoagulants
✔ Significantly increased PCR sensitivity and success rates even with very demanding applications e.g., GC-rich templates
✔ High speed and specificity make this kit suitable for multiplex PCR and high-throughput genotyping
✔ The kit’s advanced formulation permits fast cycling conditions, without compromising PCR specificity or yield

MyTaq™ Blood-PCR Kit Application Note

MyTaq™ Extract-PCR Kit

This kit is particularly suitable for solid tissues such as mouse-tail snips or ear punches. The MyTaq™ Extract-PCR Kit allows you to bypass lengthy DNA extraction protocols by providing a rapid single-tube approach for DNA extraction and amplification. The single-tube format and lack of washing steps greatly reduces the risk of sample loss and contamination that may occur during standard genomic DNA extraction procedures.

Applications

✔ High-throughput genotyping
✔ Transgene detection
✔ Knockout analysis

Advantages of MyTaq™ Extract-PCR Kit

✔ PCR is performed in proprietary buffer system using MyTaq HS Red Mix, the latest generation of very high-performance polymerases unique to Bioline
✔ Non-specific amplification is reduced using antibody hot-start technology
✔ Advanced formulation of MyTaq HS Red Mix permits fast cycling conditions, without compromising PCR specificity or yield
✔ Single-tube format significantly reduces risk of contamination in high throughout applications
✔ Amplification of any target sequence up to 5 kb
✔ Mastermix contains red dye for easy pipetting and direct gel loading

MyTaq™ Extract-PCR Kit Application Note

Try It for Yourself

If you haven’t already tried Direct PCR, why not get in touch with us at Nordic BioSite to arrange sampling of Bioline’s kits? We are also happy to receive enquiries about PCR applications not mentioned here. We look forward to hearing from you!

Contact email: info@nordicbiosite.com.