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Technical Article: The Future of High Fidelity Application Notes: Premix Ex Taq (Perfect Real Time) pCold TF Vector DICE Thermocycler and Ex Taq FAQ Mailbox Featured Products New Products Announcements
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PCR has become a basic laboratory procedure, being performed thousands of time each day in laboratories worldwide. Taq Polymerase was the first thermostable polymerase to be made available to researchers, and is still the most widely-used PCR enzyme. It is a highly processive enzyme, suitable for many routine PCR applications. However, Taqs performance is not adequate for other more demanding PCR applications, such as high-fidelity PCR, high-sensitivity PCR, or the synthesis of long or complex DNA targets. Importance of High Fidelity High polymerase fidelity (i.e. a low rate of base misincorporations, or errors) is most important in PCR applications where downstream sequencing or gene expression of the amplified product is desired. It is also significant in applications requiring amplification of low-copy-number templates (requiring many rounds of amplification), longer target sequences, or amplification and rare transcripts or allelic mutants. cDNA library construction, site directed mutagenesis, and mutation detection are also particularly sensitive to error rate. Enzyme fidelity can by influenced by a variety of factors, including template sequence (i.e. GC-rich templates generally increased error rates), cycling parameters, and reaction conditions (i.e. pH, Mg2+, dNTP concentration). However, in controlled studies, polymerases exhibit characteristic rates of base misincorporations, rates of extension from those misincorporations, and 3' 5' exonuclease or proofreading activity. These factors together result in an intrinsic error rate for each polymerase.
Polymerase Fidelity
Taq polymerase and related Thermus family polymerases generally possess a high rate of base misincorporations, a low rate of extension from these misincorporations, and lack a 3' 5' exonuclease or proofreading function. Their error rates are the highest among the most widely-studied viral and bacterial polymerases. Additionally, the low extension rate actually acts somewhat as a de facto proofreading function, as incorrect templates fall out of the amplifiable pool. However, this results in lower yield and sensitivity, particularly on longer products.
Using conventional mutant-based fidelity assays, the recorded error rates of about 10-4 are common for Taq. This number may seem low, but this means that after one fairly typical 106 fold PCR amplification of a 200 bp target, up to 33% of the resulting products may contain errors.
The expected fraction of PCRinduced mutants can be calculated according to the following formula:
Technical Article
Mixing a proofreading polymerase with Taq polymerase has been shown to increase amplification performance, and is the basis for several widelyused enzymes, including Takara Ex Taq and LA Taq. These blends provide superior amplification efficiency and product length as compared to Taq or the proofreader alone. Fidelity is also much improved over Taq Polymerase alone, but may still cause problems in some applications. Calculated Error Rate Error rate and fidelity are calculated via the following formulas: Error Rate= # misincorporated bases/# bases synthesized Fidelity= 1/error rate Most quoted error rates are experimentally based on indirect phenotypic measurements of mutant frequency, and vary widely. For example, one common method calculates the frequency of observed mutants by identifying the number of phenotypically altered colonies following bacterial transformation with a PCR-amplified DNA fragment. However, lethal amino acid substitutions derived from misincorporation of one or more incorrect bases during the PCR reaction will go unnoticed and uncounted, as they result in cell death. Fidelity rates calculated via this method are also subject to an additional level of inaccuracy because some nucleotide changes will not result in clear phenotypic changes of the expressed protein (usually beta-galactosidase). Therefore, these conventional methods of calculating error rates can provide useful comparisons within a single set of reaction conditions, but the actual results may vary widely from expected numbers.
Pyrococcus sp. polymerases (also called proofreading polymerases) have an even higher initial misincorporation rate than Taq, but because they contain a 3' to 5' exonuclease activity, they generally possess much lower error rates than Taq Polymerase or other Thermus-family polymerases. However, these enzymes often display low processivity, resulting in low product yield, reduced product length, and difficulties in optimization.
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References:
(1) Cha, R.; Thilly, W. in PCR Primer, A Laboratory Manual, 1995, 34-51. (2) Keohavong, P.; and Thilly, W. Proc. Natl. Acad. Sci. USA, 1989, 86:9253-9257. (3) Pavlov, R.; et. al. TRENDS in Biotechnology, 2004, 22:254-261. (4) Barnes, W. Proc. Natl. Acad. USA, 1994, 91:2216-220.
Technical Article
Fidelity comparison with competitors sequencing results showed only 12/249,941 mismatched bases in DNA fragments amplified using PrimeSTAR HS.
PrimeSTAR HS Offers: High Accuracy: A strong exonuclease activity, resulting in an extremely low error rate of only 12 errors per 250,000 bp. High Efficiency: Amplification efficiency higher than Taq Polymerase; excellent performance even on GC-rich templates. Robust Amplification: Tolerance to varying reaction conditions means a single PCR cycling protocol can be used to amplify products of varying sizes. Extended Product Length: Amplify targets of up to 8.5 kb on human genomic DNA; 10 kb on E. coli genomic DNA; and 22 kb on lambda DNA. Fast Reaction Times: Increased priming efficiency results in rapid annealing times and improved specificity. High Specificity: Antibody-mediated hot-start prevents false initiation events during reaction assembly due to mispriming or primer digestion.
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Comparison of Takaras New Premix Ex Taq (Perfect Real Time) versus Competing qPCR Kits
Introduction Real Time PCR offers a quantitative method to study product amounts during the early stages of a PCR reaction, when the amount of product corresponds to the amount of initial template present in the reaction. Premix Ex Taq (Perfect Real Time) is a 2X premix, specially designed for high-speed, high sensitivity real time PCR using either detection probes (i.e. TaqMan and other molecular probe technologies) or SYBR Green I (not included). This premix combines high-performance Takara Ex Taq Hot Start DNA Polymerase, which uses antibody-mediated Hot Start technology to prevent non-specific amplification, with an optimized real time PCR buffer formulation which provides increased amplification efficiency and further improved specificity for high speed real time PCR. Premix Ex Taq reactions are fast and easy, and generate exceptional PCR results on all major realtime instruments. The amplification curves generated with Premix Ex Taq demonstrate excellent sensitivity and uniformity. With the ABI Kit, only one curve appears, indicating sensitivity several logs lower than Takara s. Both the Invitrogen and Qiagen Kits resulted in amplification curves which are shifted to the right, indicating lower amplification efficiency than the Takara Kit. The curves lack uniformity when compared to Takaras Kit, indicating lower reproducibility. Additionally, reactions with Takara Premix Ex Taq can be completed in as little as 50 minutes, making it compatible with fast PCR systems. In summary, Takara Premix Ex Taq provides superior efficiency and reproducibility as compared to other leading qPCR kits.
Application Note
Features of Premix Ex Taq (Perfect Real Time) Include: Materials and Methods qPCR amplification of the human ACTB gene was performed using cDNA templates corresponding to 100 ng-1 pg of total RNA using TaqMan probes (ABI) and Premix Ex Taq (Perfect Real Time). These results were compared against results generated using three different leading competitor qPCR kits on the Roche LightCycler. Reactions were performed according to the manufacturers protocols.
Fast: Reaction can be completed in as little as 50
minutes.
High Sensitivity: Detects as few as 10 template
copies.
Versatility: Compatible with Smart Cycler,
LightCycler, ABI PRISM 7000/7700/7900 HT, Applied Biosystems 7500 Real-Time PCR Systems, and other real time PCR instruments.
Wide Dynamic Range: Possesses a dynamic
Results and Discussion Amplification efficiency was determined using Premix Ex Taq (Perfect Real Time) and three leading competitor qPCR enzymes on the Roche LightCycler. The results can be seen in the figure on the next page.
Ordering Information
Product No. TAK RR039A TAK RR039B TAK RR041A TAK RR041B TAK RR031A TAK RR031B Product Name Premix Ex Taq (Perfect Real Time) DNA Polymerase Premix Ex Taq (Perfect Real Time) DNA Polymerase SYBR Premix Ex Taq (Perfect Real Time) SYBR Premix Ex Taq (Perfect Real Time) Ex Taq R-PCR, Version 2.1 Ex Taq R-PCR, Version 2.1 Quantity 200 reactions 400 reactions 200 reactions 400 reactions 250 U 1000 U Price $219 $437 $273 $534 $189 $666
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For additional information about Takaras qPCR products see pages 12-13
Application Note
Figure 1:
Performance of SYBR Premix Ex Taq (Perfect Real Time) vs. ABIs TaqMan Universal PCR Master Mix using a Roche LightCycler. Cycling conditions: 95C, 2 min} 1 cycle 94C, 15 sec. 55C, 1 min. 72C,1 min. 45 cycles
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The pCold TF Protein Expression System Produces Soluble, Active Protein in E. coli
Elucidation of protein structure and function maintains an important role in post-genomic sequencing and analysis studies. An efficient protein production system is critical for obtaining large amounts of correctly folded recombinant protein for study. E. coli expression systems are used extensively for production of recombinant proteins, and have two major advantages over other expression systems: (1) ease of use, and (2) low cost. However, some recombinant proteins do not fold correctly during expression in E. coli, resulting in deposits of inactive insoluble protein termed "inclusion bodies". a 5' untranslated region (5' UTR), a translation enhancing element (TEE), a His-Tag sequence, and a multicloning site (MCS). A lac operator is inserted downstream of the cspA promoter to ensure strict regulation of expression. Additionally, recognition sites for HRV 3C Protease, Thrombin, and Factor Xa are located between the TF-Tag and the multiple cloning site (MCS) and function to facilitate tag removal from the expressed fusion protein. Most E. coli strains can serve as expression hosts. pCold TF DNA Vector combines high-yield cold shock expression technology with Trigger Factor (chaperone) expression in a single vector to facilitate correct protein folding, thus enabling efficient soluble protein production for otherwise intractable target proteins. The following experiment compares results generated using pCold I; pCold I co-expressed with a Chaperone Plasmid; pCold TF; and T7 promoter constructs to express various proteins.
Application Note
Series of pCold Vectors In collaboration with Prof. Masayori Inouye (University of Medicine and Dentistry of New Jersey), Takara Bio has developed the pCold DNA Vectors, a series of novel protein expression vectors. The pCold Vectors provide increased in vivo protein yield, purity, and solubility of expressed recombinant proteins using "cold shock" technology. More specifically, the cspA (cold shock protein A) promoter and related elements have been incorporated into these vectors to upregulate target protein production at lowered incubation temperatures (37C-15C). This temperature drop also suppresses expression of other cellular proteins, represses protease activity, and temporarily halts overall cell growth. This process allows expression of target proteins at high yield, high purity (up to 60% of cellular protein), and increased solubility as compared with conventional E. coli expression systems. Co-expression of one or more chaperone proteins during expression of a heterologous target protein has proven effective for obtaining increased amounts of soluble recombinant protein (see Takara's Chaperone Plasmid Set (TAK 3340)). This procedure, though, lacks the convenience of a single transformation step.
Materials and Methods pCold DNA I and pCold TF DNA cloning and expression procedures* were conducted as follows: 1) Insert the target gene to the multicloning site of the pCold DNA vector for expression. 2) Transform the E.coli host strain (e.g. BL21) with the expression plasmid, and select for ampr transformants. 3) Inoculate the transformants into medium including 50 g/ml of ampicillin, and culture with shaking at 37C. 4) At OD600= 0.4 - 0.5, refrigerate the culture at 15C (without shaking) for 30 minutes. 5) Add IPTG to a final concentration of 0.1- 1.0 mM, and continue the culture with shaking at 15C for 24 hours. 6) Collect the cells, and confirm the expression of the target protein with SDS-PAGE in soluble and insoluble fractions or activity assay. Expression from T7 promoter-driven vectors was performed using a standard protocol utilizing IPTG induction and subsequent culturing at 37C. *Cultivation/induction conditions (culture medium, aeration, timing of induction, concentration of inducer, cultivation time after induction) should be optimized for each target protein. 6
pCold TF Vectors Takara's pCold TF DNA Vector is a fusion cold shock expression vector that expresses a molecular chaperone (Trigger Factor (TF)) as a soluble tag. Trigger Factor is a prokaryotic ribosome-associated chaperone protein (48 kDa) which facilitates co-translational folding of newly expressed polypeptides. Because of its E. coli origin, TF is highly expressed in E. coli expression systems. The pCold TF DNA Vector consists of the cspA (cold shock) promoter plus additional downstream sequences including
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kDa
pCold TF 1 2
pCold 1 2
pCold + Chaperone 1 2
T7 1 2 1. Cell extract solution 2. Soluble fraction target protein co-expressed trigger factor
kDa 97 66 45 31 22
Trx
GST
Nus
1 2 3 1 2 3 1 2 3
Application Note
97 66 45 31 22
Figure 2: Increased Expression of Soluble Protein B Using the pCold Expression System
Results and Discussion Protein expression using the pCold TF Expression Vector was compared with protein expression using (1) the pCold DNA I Vector alone, (2) co-expression using the pCold DNA I Vector with Takara's Chaperone Plasmid pTf16, and (3) a T7 promoter expression system which included other tags for solubilization. Figure 1 demonstrates the successful production of enzyme protein A using the pCold TF system. Expression of this protein, with an estimated molecular weight of 29 kDa, was not seen as an exact band with either the T7 expression system or even with pCold I (either individual expression or chaperone co-expression). However, the expression of the target protein and target plus tag (29 kDa and 52 kDa) was observed using pCold TF, and most of the obtained protein was in soluble form. Subsequent assays confirmed that the expressed enzyme A retains activity even as a fusion protein. Figure 2 demonstrates improved levels of soluble protein B using pCold TF. Expression of soluble enzyme protein B (M.W: ~63 kDa) was not observed using either pCold DNA I alone or pCold I co-expressed with chaperone proteins, nor with a T7 expression vector that included other tags for solubilization (Trx Tag [~12 kDa], Nus Tag [~55 kDa], and GST Tag [~26 kDa]). However, when the pCold TF DNA Vector was used, the target protein was present at an expression level much higher than with other systems and tags, and most of the expressed target protein was observed in the soluble fraction. (Note: The molecular weight of the target protein is larger than its actual size and varies due to fused expression with different tags). In summary, the pCold TF expression system offers a convenient high yield, high purity alternative for efficient soluble protein expression of otherwise intractable target proteins.
Ordering Information
Catalog No. TAK 3365 Product Name pCold TF Vector Quantity 25 g Price $727
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FAQ Mailbox
PrimeSTAR HS DNA Polymerase FAQ
Can PrimeSTAR HS reactions use the same PCR cycling conditions that are used with Taq Polymerase?
PrimeSTAR HS cannot use the same PCR cycling conditions that are used with Taq Polymerase. Takara strongly recommends following the conditions described in the PrimeSTAR HS product protocol, since the characteristics of this enzyme are very different from those of Taq Polymerase. Takara recommends the following initial cycle protocol for primers with a Tm of >55C: denaturing step, 98C, 10 sec; annealing step 55C, 5 sec.; extension step, 72C, 1 min./kb; for 30 cycles.
FAQ Mailbox
What advantage is offered by Takaras measurement of PrimeSTAR HSs fidelity using sequence analysis?
Simple comparison of the fidelity rates available for different PCR enzymes is not possible due to the multitude of different fidelity measurement techniques employed by different manufacturers. Takara Bio has determined PrimeSTARs error rate based upon genotype, that is, the error rate as determined by actual sequence analysis. The method Takara Bio used to obtain their fidelity data is presented below: Eight arbitrarily selected GC-rich regions were amplified with PrimeSTAR HS and other enzymes, using the Thermus thermophilus HB8 genomic DNA as a template. Each PCR product (approx. 500 bp each) was cloned into a suitable plasmid. For each different DNA region cloned, multiple clones were picked, and subjected to sequence analysis. Sequence analysis results of DNA fragments amplified using PrimeSTAR HS demonstrated only 12 mismatched bases per 249,941 total bases. This data confirms PrimeSTAR HSs extremely high fidelity, with a calculated error frequency of only 0.0048%.
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FAQ Mailbox
What is the purpose of the ROX reference dye included with the SYBR Premix Ex Taq?
ROX (i.e. Carboxy-X-Rhodamine) is a convenient internal reference standard for use in normalizing signals due to non-PCR related fluorescence fluctuations that occur either between wells or over time. Please note that two types of ROX Reference Dye (Original Version ROX and ROX II) are supplied with this product. Use the Original Version ROX for normalization with the ABI PRISM 7000/7700/7900HT and Applied Biosystems 7300 Real-Time PCR Systems. Use ROX II reference dye for normalization whith Applied biosystems 7500 Real-Time PCR System.
How do I determine the number of qPCR reactions for my experiments? For example, if I have two different cell lines and want to characterize three different genes in each?
For each of the 3 genes, a standard curve (e.g. composed of 7 data points) plus 2 experimental samples that are run in triplicate are performed. Therefore, 3(triplicate) x (7 pts + 2 samples) x 3(genes) = 81 reactions are required for 3 genes. One package of SYBR Premix Ex Taq (Perfect Real Time) contains sufficient reagent for 200 reactions (50 l reaction volume).
Can you make a master mix of the ROX reference dyes and SYBR Premix Ex Taq to help avoid pipetting errors?
Original Version ROX Reference Dye can be premixed. Add 40 l of ROX to 1 ml of SYBR Premix Ex Taq. Store this solution at 4C (protected from light), and use within one month for best performance. ROX Reference Dye II should not be premixed prior to reaction assembly.
Can SYBR Green I dye precipitate? Is there a good way to resuspend SYBR Green I?
A greenish-yellow precipitate can sometimes be observed in SYBR Premix Ex Taq when stored at -20C. If this occurs, dissolve the precipitate completely by letting the tube stand at room temperature for sev-
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Highly Efficient Gradient PCR of an 8 kb DNA Fragment using Takara Ex Taq DNA Polymerase and the Takara PCR Dice Thermocycler
Introduction:
Takaras Ex Taq DNA Polymerase is a polymerase mix which offers high amplification efficiency (up to 100X Taq), superior reproducibility, and extended product length (to 30 kb on DNA). In both protocols, robust, reproducible amplification was apparent. These results demonstrate that Takaras Ex Taq DNA Polymerase and Dice Thermocycler can be combined to produce easy optimization and superior reproducibility and yield even with challenging DNA fragment lengths.
M 52C 72C M
Application Note
The Takara Thermocycler Dice provides convenience, reliability, multifunctionality, and a gradient feature to select temperature conditions at each step, and program a temperature gradient up to 20C across the rows of a sample block. This allows optimization of reaction conditions in a single experiment. The following experiment demonstrates optimization of the PCR conditions for an 8 kb DNA fragment amplified with Ex Taq DNA Polymerase on the PCR Thermocycler Dice.
Figure 2: Evaluation of annealing temperatures in three-step gradient PCR. Higher annealing temperatures reduced the amplification of non-specific products.
Ordering Information
Product No. TAK RR001A TAK RR001B TAK RR001C Product Name Ex Taq DNA Polymerase Ex Taq DNA Polymerase Ex Taq DNA Polymerase Quantity 250U 1000U 3000U Price $158 $567 $1483
To order the Takara Thermocycler Dice (TP600) please contact Sanyo Biomedical at 905-760-4049 (Canada) or Fisher Scientific at 800-766-7000 (USA).
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Featured Products
SYBR Premix Ex Taq (Perfect Real Time)
For superior real time PCR using SYBR Green I
Features
High sensitivity: Detects as few as 100 copies. Versatility: Compatible with most qPCR instruments.
Amplification Curve (upper panel) and Melting Curve (lower panel) for SYBR Premix Ex Taq (Perfect Real Time) on the SmartCycler (Cepheid).
Featured Products
Wide dynamic range: Possesses a dynamic range of 7-8 orders of magnitude ( DNA template). High speed capability: Optimized buffer system allows quick reaction times. Convenient: Separate tubes of ROX reference dye are supplied. SYBR Premix Ex Taq (Perfect Real Time) is a convenient (2X) premix consisting of Takara's high sensitivity, high efficiency Ex Taq Hot Start DNA Polymerase, SYBR Green I, dNTPs, and an optimized real time buffer which provides superior specificity and increased amplification efficiency for real time PCR, plus separate tubes of ROX Reference Dye. Antibody-mediated hot start technology is used for reduced background.
Ordering Information
Catalog No. TAK RR041A TAK RR041B Product Name SYBR Premix Ex Taq SYBR Premix Ex Taq Quantity 200 reactions 400 reactions Price $273 $534
Amplification curve for Premix Ex Taq (Perfect Real Time) using the TaqMan Gene Expression Assay on the Applied Biosystems 7500 Real-Time PCR System.
Ordering Information
Catalog No. TAK RR039A TAK RR039B Product Name Premix Ex Taq Premix Ex Taq Quantity 200 reactions 400 reactions Price $219 $437
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Ex Taq R-PCR Version 2.1 offers the excellent specificity, sensitivity, and low background required for superior real time PCR.
Featured Products
Ex Taq R-PCR Version 2.1 contains a monoclonal antibody to Taq Polymerase, which binds to the polymerase until the temperature is elevated. The binding of this antibody prevents nonspecific amplification due to mispriming and/or formation of primer dimers during reaction assembly. An optimized buffer formulation provides increased specificity for real time PCR, further enhancing performance.
Ordering Information
Catalog No. TAK RR031A TAK RR031B Product Name Ex Taq R-PCR, Ver 2.1 Ex Taq R-PCR, Ver 2.1 Quantity 250 Units 1000 Units Price $189 $666
nomalize the fluorescent signal between reactions (for instruments that are equipped with this option).
Melting curve.
Detection of Rat Atp5fl with the II System. A real time One-Step RT-PCR reaction was performed using total RNA (1 pg100 ng) prepared from rat liver as a template. The target gene was detected in the range of 10 pg100 ng. The melting curve shows that a single product was amplified at all template concentrations.
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Catalog No. TAK RR026 Product Name Real Time One Step RNA PCR Kit Quantity 100 reactions Price $309
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New Products
pCold TF Vector
For increased soluble protein expression
Features
High yield recombinant protein: Up to 60%, of expressed intracellular protein is the target protein. Soluble expression level is increased: Due to fused expression with Trigger Factor, proteins that are insoluble in conventional expression systems may be expressed in soluble form. Wide range of E. coli hosts: Compatible with most E. coli strains. More protease are available for tag release: Cut sites for 3 proteases are included.
Amp
M
cspA 3UTR Multiple cloning site Factor Xa site Thrombin site HRV 3C Protease site Trigger Factor (TF) His Tag TEE cspA 5UTR lac operator cspA promoter
New Products
IG 13
pCold TF DNA
lac I
Quantity 25 ug
(5,769 bp)
Takara's pCold TF DNA Vector is a fusion cold shock expression vector that expresses Trigger Factor (TF) chaperone as a soluble tag. Trigger Factor is a prokaryotic ribosome-associated chaperone protein (48 kDa) which facilitates co-translational folding of newly expressed polypeptides. Because of its E. coli origin, TF is highly expressed in E. coli expression systems. The pCold TF DNA Vector consists of the cspA promoter plus additional downstream sequences including a 5' untranslated region (5' UTR), a translation enhancing element (TEE), a His-Tag sequence, and a multicloning site (MCS). A lac operator is inserted downstream of the cspA promoter to ensure strict regulation of expression.
ColE1 ori
Ordering Information
Catalog No. TAK 3365 Product Name pCold TF Price $767
Ordering Information
Catalog No. Product Name Quantity 1 set TAK 3367 SPP System I Please visit our website for additional products. Price $720
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Try it today and prove it to yourself. Call or visit our web site to request a sample of this product.
PrimeSTAR HS DNA Polymerase is a novel new high fidelity PCR enzyme which provides maximum fidelity as well as extended product length (8.5 kb for human genomic DNA; 22 kb for DNA). Targeted for demanding cloning (i.e. amplification of cDNA libraries) and sequencing applications, it offers extremely high accuracy, and fidelity calculated by sequence analysis. It also offers excellent amplification efficiency and shortened reaction times. Finally, the antibody-mediated hot start formulation prevents false initiation events during reaction assembly due to mispriming or primer digestion, thus, lowering background.
New Products
- 2kb
Comparison of PrimeSTARTM HS Amplification Efficiency with Competitors on a 2 kb Human Genomic DNA Fragment. Superior amplification efficiency was apparent using PrimeSTARTM HS on a human genomic (DCLRE1A) 2 kb template. Human genomic DNA was used in the following quantities: Lane1: 0 ng (dH2O), Lane 2: 100 pg, Lane 3: 1 ng, Lane 4: 10 ng, Lane 5: 100 ng.
M1 M2 1
10 M2 M1
1. DCLRE1A 4 Kb 2. beta-globin 8.5 Kb 3. beta-globin 6 Kb 4. DCLRE1A 2 Kb 5. p53 0.5 Kb 6. p53 4 Kb 7. beta-globin 7.5 Kb 8. DCLRE1A 8 Kb 9. DCLRE1A 1 Kb 10. p53 6 Kb M1: -Hind III-digest M2: pHY Marker
Ordering Information
Catalog No. TAK R010A TAK R010B Product Name PrimeSTAR HS PrimeSTAR HS Quantity 250 Units 1000 Units Price $218 $785
A Single PCR Cycling Protocol Produces Optimal Amplifcation of Fragments of Varying Sizes. A single protocol allows efficient amplification of target fragments from 0.5 - 8.5 kb, making PrimeSTAR HS an excellent choice for a cDNA library cloning.
Ex Taq, LA Taq, SPP System and PrimeSTAR are trademarks of, and RetroNectin is a registered trademark of Takara Bio Inc. SYBR is a registered trademark of Molecular Probes, Inc. TaqMan is registered trademark of Roche Molecular Systems. LightCycler is a trademark of the Roche group. Platinum is registered trademark of Invitrogen. Smart Cycler is a registered trademark of Cepheid. PRISM is a registered trademark of Applied Biosystems. MJ Opticon is a registered trademark of BioRad Laboratories, Inc. Takara PCR Related Products are sold under a licensing arrangement with Roche Molecular Systems and F. Hoffman La Roche Ltd.and Applied Biosystems. Takara Bios Hot-Start PCR-Related products are licensed under U.S. Patent 5,338,671 and 5,587,287 and corresponding patents in other countries. His Tag sequences in pCold I, II, and TF are licensed from Hoffman-LaRoche, Inc. pCold DNA and the SPP System are covered by U.S. patents and pending patents owned and issued by the University of Medicine and Dentistry of New Jersey.
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