Bio-pass recommendation: Phusion high-fidelity polymerase in the fighter

Abstract: PCR technology can be described as the field of molecular biology one of the greatest technological innovation, PCR amplification of PCR technology is the key enzyme core. Taq enzyme cheap and efficient, but the amplification of short length, error rate, the consequences could be serious; high-fidelity enzyme error rate is small, but the amplification efficiency of low prices and expensive. Hybrid enzyme which is not to please. Recommended new biological pass Phusion ® DNA polymerase, amplification efficiency is higher than the Taq 1 times, high yield; fidelity than Taq 50 times higher, much higher than the famous high-fidelity Pfu enzyme, with less time is short, high yield, bright bands, the most high-fidelity, cheaper, how can you not try! Sony VGP-BPL8 Battery
If the PCR product cloning and expression of the need for subsequent research, the need for high-fidelity PCR to reduce PCR errors caused by the process of mutation, we tend to choose high-fidelity DNA polymerase. General high-fidelity DNA polymerase because of their own validation function determines that low efficiency, low yield, weak bands, time slow. The famous high-fidelity enzyme Pfu Taq enzyme efficiency of only about 1 / / 5. People often lament, "less expensive and easy to use (refer to yield low)"! In order to improve efficiency, some company has launched a hybrid enzyme, with the parity function with the Taq DNA polymerase mix, moderation is not as efficient as the result of Taq polymerase, fidelity as Pfu, Taq generally three times higher than the price also not cheap. Sony VGP-BPL9 Battery
Finnzymes Corporation (now Thermo Fisher Scientific acquired) Off the beaten track, the double-stranded DNA binding protein (purple) and a new class of heat with a proofreading function of bacterial enzyme (green) fused to form a unique high performance polymerase - Phusion ® DNA polymerase. This time, two issues have been resolved. Sony VGP-BPL13 BatteryFigure 1. Phusion DNA polymerase of unique structure

Quick step
Sony VGP-BPL18 BatteryBecause of this unique blend of technology, Phusion DNA polymerase DNA synthesis is the speed and accuracy in any previous single enzyme are difficult to match even the most complex template can be accurately and quickly complete the reaction. Compared with other enzyme, Phusion DNA polymerase binding to DNA in each reaction are able to more nucleotide incorporation, its high processing capability greatly reduces the extension of time steps, thus accelerating the entire reaction. It is an extension of the rate of Pfu enzyme is 10 times faster than the Taq enzyme doubled to 15-30 sec / kb. If you want faster, you can choose high-fidelity polymerase Phusion ® Flash fast, an extension of time for each kb base only 15 seconds or less, the shortest reaction time, enzyme dosage was significantly lower than other common polymerase. Phusion DNA polymerase high-speed and high efficiency make it get a lot of product in the shortest time.
Sony VGP-BPL21 BatteryFinnzymes have done experiments, from Thermus sp. Genomic library in 16 randomly selected clones, and with three different heat-resistant DNA polymerase amplification. The results showed that, Phusion ® DNA polymerase with strong performance and speed, it can be successfully amplified 16 randomly selected fragments of 15 (94%), and high yield. The Pfu DNA polymerase success rate was 56%, Taq DNA polymerase success rate was 62%, both significantly lower than the output of Phusion ® DNA polymerase. And can be seen from the figure, Phusion ® DNA polymerase extension of the time and volume was significantly lower than with the enzyme Pfu enzyme with Taq quite, but higher yields. Such a strong performance second to none. Sony VGP-BPS8 Battery
Figure 2. Phusion DNA polymerase set of ultra-high fidelity, fast and strong performance in one
Less error-prone
Sony VGP-BPS9 BatteryMore importantly, Phusion DNA polymerase fidelity on the market today is the highest thermal stability of the polymerase. LacI using improved methods of detection, Phusion DNA polymerase error rate than Taq DNA polymerase with a low around 50 times lower than six times Pfu DNA polymerase, is ideal for high fidelity PCR. For the study of gene function, need to engage in site-directed mutagenesis, gene cloning and expression analysis work for, Phusion DNA Polymerase is undoubtedly the better choice. Sony VGP-BPS9/S Battery
Taq polymerase error probability is higher, and the average length of no more than amplified 3.5kb. The Phusion DNA polymerase amplification may be more than 10kb (<20kb) the long fragment, fidelity is not easy to make mistakes, coupled with high-fidelity PCR enzyme is much faster rate than normal, so you can use the Phusion DNA polymerase effectively save a lot of time. Sony VGP-BPS9/B BatterySave time, save the amount of Because of its unique structure, Phusion DNA polymerase is very efficient. In the PCR reaction, the enzyme dosage was significantly lower than other common polymerase. Researchers have used three different DNA polymerase amplification of a length of 3.8kb of the human β globin gene. Using Phusion DNA polymerase can not only successfully amplified the 3.8kb genomic fragment, and annealing and extension steps in 1 minute, significantly faster than the other two polymerases. A unit of Phusion DNA polymerase to obtain the product of more than 5 units of Pfu DNA polymerase yields, improved more than 2.5 units of Pfu yields are much higher.
Figure 3, with less of the enzyme - the product of more Sony VGP-BPS9A/B Battery
Applicability

In addition, Phusion DNA polymerase can tolerate a variety of PCR inhibitors maximum, so only a few optimization steps can be very efficient amplification. This feature has also spawned an unprecedented direct PCR. For blood, tissue and other samples, even FFPE samples, without extracting DNA, high-fidelity PCR amplification can be carried out directly. Sony VGP-BPS9A Battery
Phusion DNA polymerase have multiple versions to choose from. In addition to the above-mentioned Phusion Flash, there Phusion hot start high-fidelity DNA polymerase II, its highly specific and strong aggregation, determines that it is ideal for hot start PCR. Characteristics of several enzymes and products in the form below: Sony VGP-BPS10 Battery Phusion high-fidelity DNA polymerase
Phusion hot start high-fidelity DNA polymerase II
Phusion Flash
High-fidelity DNA polymerase
Phire Hot Start Polymerase IIDNA

Enzyme
Feature
Flat end or 3 'A-side
Plain End
Plain End
Plain End
Plain End

Target fragment length
≤ 20 kb
≤ 20 kb
≤ 20 kb
≤ 7.5 kb

Hot Start
No
Be
Be
Be

Recommended an extension of time
15-30 s / kb
15-30 s / kb
15 s / kb
10-15 s / kb

Correction
Be
Be
Be
General

Fidelity vs. Taq
52x
52x
25x
2x
Product
Form
Single enzyme 1
Master Mix2
Kit 3
Cheap and wide choice
Sony VGP-BPS13 BatteryPhusion DNA polymerase performance so strong, and the price is quite close to the people. With Phusion high-fidelity PCR kit (except the template with primers, all other reagents required), for example, list price for just 555 yuan to 125 responses (each 20μl), the cost of a single response was 4.44 yuan, 1878 yuan or 500 response may have simply too much to force it! This is also the editor of Biological highly recommended through one of the reasons. Sony VGP-BPS14 BatteryPhusion DNA polymerase (containing enzyme, buffer, Mg ions, DMSO) as long as 879 yuan / 100U, enough to make 100 50ul reaction. To know PfuUltra catalog price 100U single enzyme (40 responses) also more than 1900 ah!
Sony VGP-BPS18 BatteryPhusion DNA polymerase offers a variety of other options, a single hot-start high-fidelity enzyme (1067 yuan / 100U), also for GC-rich templates, or especially with HF buffer premixed synthase MasterMix products.
Sony VGP-BPS21 BatteryFor site-directed mutagenesis needs to be done for gene function analysis of the students, Thermo Fisher has launched configuration super high-fidelity enzyme Phusion DNA polymerase 20-directed mutagenesis kit, the price as long as 1195,20 second reaction! Done this experiment the students must be very clear that this experiment is the key to a DpnI restriction endonuclease and amplified by high fidelity PCR enzyme, another brand of this kit had better response to 4590 dollars 10 times, now Thermo Fisher this price, hey, only about 1 / 4, 20 reactions (with 10 control reactions), reaction times more than doubled the performance of high-fidelity enzymes can be more powerful it! However, the lack of competent cells is estimated. Sony VGP-BPS22 Battery