Return to site

5 Features to Look for When Comparing Different Types of Bst Polymerases

Amplification speed, reverse transcription activity, impurity tolerance, specificity, and what

August 22, 2022

When it comes to finding the best Bst Polymerase for your research, it’s tempting to look at the polymerase with the highest amplification speed, reverse transcription activity, impurity tolerance, and specificity. But inevitably, cost becomes a factor. Researchers have to consider which options are most affordable while meeting the needs.

Today’s midrange Bst Polymerases like Bst DNA Polymerase offer great performance. Compared with wild-type Bst DNA polymerase (large fragment), Bst DNA Polymerase has been greatly improved in terms of amplification speed, yield, salt tolerance, and thermal stability. Therefore, it is an excellent enzyme for most isothermal amplification.

But how do you know which affordable Bst Polymerase is best for you and your research? As you search for the right fit, look out for these 5 must-know features that will have the greatest impact on your productivity while staying within budget:

 

Amplification Speed

Amplification speed is very important as it can help to shorten the experimental time, resulting in higher research efficiency. The wild-type Bst DNA polymerase (large fragment) has a comparatively low amplification speed, so the detection time takes long time. Fortunately, with electronic reconstruction and enzyme evolution technology, the performance can be largely improved, including the amplification speed.

 

Reverse Transcription Activity

Reverse transcription activity is extremely important when you are using RNA templates. The wild-type Bst DNA polymerase (large fragment) and Bst DNA Polymerase have very limited reverse transcription activity, so they are not suitable for the RNA samples. With genetic engineering, Bst Polymerase has strong reverse transcription activity and can carry out reverse transcription without any other enzymes. Bst DNA/RNA Polymerase is a mixture of Bst Polymerase and extremely thermostable reverse transcriptase (65°C tolerant). With addition of reverse transcriptase, it is even more suitable for the isothermal amplification reaction of RNA. Low-sensitivity RNA molecules can be detected with Bst DNA/RNA Polymerase.

 

Impurity Tolerance

The impurities in samples can greatly impact the amplification of the enzymes. Without optimization, wild-type Bst DNA polymerase (large fragment) has poor performance when there are contaminations in the sample, resulting in the failure of the detection. Fortunately, with electronic reconstruction and enzyme evolution technology, the impurity tolerance can be largely improved.

 

Specificity

Specificity is an important parameter to evaluate the ability of a screening test to detect a true negative or positive. With high-speed amplification characteristics, the wild-type Bst DNA polymerase (large fragment) will inevitably lead to false positive amplification, especially when primers are at high concentration, resulting in low specificity. Fortunately, with genetic engineering, higher specificity can be achieved. Besides, the engineered enzyme has a strong recognition ability to dUTP. The dTTP needed for amplification reaction can be completely replaced by dUTP, so the amplification products all contain dUTP. By adding Heat-Labile Uracil DNA Glycosylase (HL-UDG), aerosol pollutants will be completely removed in the initial reaction stage. The HL-UDG can be later inactivated irreversibly within 3 min at 65°C, which not only eliminates the pollutants but also ensures the normal amplification of nucleic acid. Therefore, the false positive caused by aerosol pollution in the reaction can be greatly reduced.

 

Whether This Enzyme is Lyophilizable

Freeze drying or lyophilization technology has been widely used in the fields of medicine, biological products, food, blood products and active substances, and has been gradually applied in enzyme reagents. The advantages of freeze drying are as follows:

  • Stable structure
  • The biological activity is basically unchanged
  • Porous, with good rehydration and good efficacy
  • Eliminates 95% ~ 99% moisture and can be stored at room temperature or in a refrigerator for a long time

However, most Bst Polymerases on the market are not ideal for lyophilization, as there are two prerequisites. First, the enzyme shall have a high concentration. Second, the enzyme has to be glycerol-free. At SBS Genetech, we provide both Bst Polymerase (glycerol-free) and Bst DNA/RNA Polymerase (glycerol-free) which are lyophilizable. Besides, we also offer Bst DNA/RNA Lyo Buffer. With this optimized buffer, there is no need to optimize the conditions of the freeze-drying protective agent.