Shop All PCR Specific Buffers

GC-Rich PCR Performance


Reaction Speed


UltraPure™ Buffer-Saturated Phenol Invitrogen™

UltraPure™ Buffer-Saturated Phenol is used in the purification of nucleic acids. The reagent, which consists of UltraPure™ Phenol that has been saturated with Tris-HCl buffer, is already buffer equilibrated to pH >7.4. When mixtures are extracted with UltraPure™ Buffer- Saturated Phenol, proteins are denatured and collect in the organic phase or at the interphase, while most nucleic acids remain in the aqueous phase. UltraPure™ Buffer-Saturated Phenol contains no preservatives. It is packaged under an inert gas in shatter-resistant, plastic-coated amber bottles.

Performance and Quality Testing: The appearance of the solution is evaluated at room temperature. No RNase or DNase activity detected.

UltraPure™ Glycerol Invitrogen™

UltraPure Glycerol is suitable for the concentration and storage of enzymes at low temperatures. A 50% (v/v) solution will not freeze at -20°C. Because it has a density of 1.26 g/mL, glycerol frequently is used as a component in electrophoresis loading buffers. UltraPure Glycerol gradients can be used to purify proteins, bacteriophage, or organelles. Redistilled UltraPure Glycerol is >=99% pure.

Performance and quality testing: no DNase, RNase, or protease activity detected

Platinum™ PCR SuperMix High Fidelity Invitrogen™

Platinum® PCR SuperMix, High Fidelity, is a ready-to-use mixture of DNA polymerase, salts, magnesium, and dNTPs for efficient PCR amplification. You need only add template and primers, reducing set-up time by half (see figure). With Platinum® PCR SuperMix, High Fidelity, you'll:

• Achieve greater than six-times higher fidelity than Taq DNA polymerase
• Amplify fragments up to 15 kb
• Produce products that are mixed blunt and 3-A' ends; however, the majority of ends will have a 3-A' overhangs
• Minimize PCR optimization

Using Platinum® PCR SuperMix, High Fidelity
Platinum® PCR SuperMix, High Fidelity, is for high-specificity, high-fidelity DNA amplification applications, such as cloning or mutagenesis. High fidelity is provided by a mixture of Taq DNA polymerase and the proofreading Pyrococcus species GB-D polymerase. High specificity is achieved by anti-Taq antibodies, allowing for automatic 'hot-start' PCR. This hot-start capability increases specificity and yield and allows for room-temperature reaction assembly.

UltraPure™ Formamide Invitrogen™

UltraPure™ Formamide is commonly used for denaturing nucleic acids for sequencing gel electrophoresis, electron microscopy, and hybridization. A liquid at room temperature, UltraPure™ Formamide is vacuum distilled and packaged under dry nitrogen and can be used as supplied for many applications within three months of date of purchase. Because the breakdown products of formamide degrade nucleic acids, for sensitive applications formamide should be deionized by treatment with a mixed-bed ion-exchange resin immediately before use.

Phenol:Chloroform:IAA, 25:24:1, pH 6.6 Invitrogen™

Ambion® Acid Phenol:Chloroform:IAA (25:24:1) is premixed and supplied at pH 6.6 (pH 7.9 ± 0.2), with included buffer. Acid Phenol:Chloroform:IAA is provided in one bottle of 400 mL. In RNA and DNA extraction procedures, Acid Phenol:Chloroform:IAA (pH 7.9) helps to stabilize the interface and prevents foaming when mixing. Preparation of phenol for use in molecular biology applications is a time-consuming and often hazardous procedure due to its toxic and corrosive nature. Ambion® premixed, quality tested, saturated phenols are ready-to-use, eliminating handling problems and offering a more convenient, safer, and easier alternative to preparing solutions from crystalline phenol.

Linear Acrylamide (5 mg/ml) (1 ml Tube) Invitrogen™

Ambion® Linear Acrylamide (5 mg/mL) is for use as a nucleic acid coprecipitant and is ideal for PCR and RT-PCR reactions since small amounts of contaminating nucleic acids present in other carriers could be amplified. It is supplied in five tubes containing 1 mL each.

• Ideal for RT-PCR
• Increases pellet mass
• Quantitative recovery of low concentrations (ng/mL) of nucleic acid
• Prevents pellet loss in nuclease protection assays

What is a Coprecipitant?
Coprecipitants are inert substances used to aid recovery of nucleic acids during alcohol precipitations. While they can be used for precipitating large amounts of nucleic acids, they are essential for quantitative recovery of small amounts of nucleic acids in dilute solutions. Often, the use of such molecules is desirable for no other reason but visualization of the pelleted precipitate after centrifugation. Linear acrylamide offers advantages over other coprecipitants in that it is chemically synthesized and not treated with reagents (e.g. proteases) derived from biological sources. Therefore, it is not contaminated with biological material making it ideal for use upstream of RT-PCR. It has been shown to precipitate picogram amounts of DNA fragments larger than 20 base pairs while failing to precipitate shorter fragments and free nucleotides. Linear acrylamide is useful for separating reaction products from unincorporated nucleotides and from most oligonucleotide primers. It should be used at a final working concentration of 10–20 µg/mL. Linear acrylamide will not interfere with A260/280 readings.

EDTA (0.5 M), pH 8.0, RNase-free Invitrogen™

Ambion® Molecular biology grade, 0.5 M EDTA, pH 8.0 solution is supplied in one bottle containing 100 mL. The solution is certified RNase-free, economical, and ready-to-use. Due to the ubiquitous presence of RNases, manufacturing products for use with RNA is especially challenging. Ambion®'s nuclease-free reagents and buffers are manufactured in facilities specifically designed to prevent the introduction of nucleases. Highly sensitive RNase assays are performed at several different stages of the manufacturing process to ensure the highest quality. These reagents are rigorously tested for contaminating nonspecific endonuclease, exonuclease, and RNase activity.

MgCl2 (1 M) Invitrogen™

Molecular biology grade, Ambion® 1 M MgCl2 solution is supplied in one bottle containing 100 mL. The solution is certified RNase-free, economical, and ready-to-use. Due to the ubiquitous presence of RNases, manufacturing products for use with RNA is especially challenging. Ambion®'s nuclease-free reagents and buffers are manufactured in facilities specifically designed to prevent the introduction of nucleases. Highly sensitive RNase assays are performed at several different stages of the manufacturing process to ensure the highest quality. These reagents are rigorously tested for contaminating nonspecific endonuclease, exonuclease, and RNase activity.

Proteinase K, recombinant Invitrogen™

Proteinase K from the fungus Engyodontium album is a nonspecific serine protease that is useful for general digestion of proteins. Proteinase K remains active:

• Over a wide pH range—optimal activity between 6.5 and 9.5
• Under denaturing conditions—e.g., in the presence of SDS or urea
• In the presence of metal chelating agents—e.g., EDTA
• At comparatively high temperatures—optimum digestion temperature is 65°C

Applications
Removal of endogenous nucleases during the preparation of DNA and RNA; preparation of tissue sections for in situ hybridization.

Performance and quality testing
Endodeoxyribonuclease and exodeoxyribonuclease assays; liquid form tested for absence of RNase activity.

Unit definition
One mAnson unit is described as that amount of enzyme that liberates 1 µmole of Folin-positive amino acid within 1 min at 37°C using hemoglobin as a substrate.

Sodium Acetate (3 M), pH 5.5, RNase-free Invitrogen™

Ambion® Molecular biology grade, 3 M Sodium Acetate solution is supplied in one bottle containing 100 mL. The solution is certified RNase-free, economical, and ready-to-use. Due to the ubiquitous presence of RNases, manufacturing products for use with RNA is especially challenging. Ambion®'s nuclease-free reagents and buffers are manufactured in facilities specifically designed to prevent the introduction of nucleases. Highly sensitive RNase assays are performed at several different stages of the manufacturing process to ensure the highest quality. These reagents are rigorously tested for contaminating nonspecific endonuclease, exonuclease, and RNase activity.

Potassium Acetate (3 M), pH 5.5, RNase-free Invitrogen™

Ambion® Molecular biology grade, 3 M Potassium Acetate, pH 5.5 solution is supplied in one bottle containing 100 mL. The solution is certified RNase-free, economical, and ready-to-use. Due to the ubiquitous presence of RNases, manufacturing products for use with RNA is especially challenging. Ambion®'s nuclease-free reagents and buffers are manufactured in facilities specifically designed to prevent the introduction of nucleases. Highly sensitive RNase assays are performed at several different stages of the manufacturing process to ensure the highest quality. These reagents are rigorously tested for contaminating nonspecific endonuclease, exonuclease, and RNase activity.

AccuPrime™ Pfx DNA Polymerase Invitrogen™

AccuPrime™ Pfx DNA Polymerase is ideal for high-fidelity amplification of DNA fragments for downstream applications such as cloning and mutagenesis. High fidelity is provided by a proprietary enzyme preparation containing recombinant DNA polymerase from Thermococcus species KOD with proofreading (3´→5´ exonuclease) activity. Platinum® anti-Pfx DNA polymerase antibodies inhibit polymerase activity, providing hot-start capabilities for improved PCR specificity (see figure). Thermostable AccuPrime™ accessory proteins enhance specific primer-template hybridization during every cycle of PCR, increasing specificity, yield, and robustness over Platinum® Pfx alone. With AccuPrime™ Pfx DNA Polymerase you will:

• Achieve greater than 26 times higher fidelity than Taq DNA polymerase
• Amplify fragments up to 12 kb
• Minimize PCR optimizationApplications
Amplification of DNA from complex genomic, viral, and plasmid templates; and RT-PCR. AccuPrime™ Pfx DNA Polymerase is especially effective where very high levels of fidelity are required.

Proteinase K Solution (20 mg/mL), RNA grade Invitrogen™

Proteinase K from the fungus Engyodontium album is a nonspecific serine protease that is useful for general digestion of proteins. Proteinase K remains active:

• Over a wide pH range—optimal activity between 6.5 and 9.5
• Under denaturing conditions—e.g., in the presence of SDS or urea
• In the presence of metal chelating agents—e.g., EDTA
• At comparatively high temperatures—optimum digestion temperature is 65°C

Applications
Removal of endogenous nucleases during the preparation of DNA and RNA; preparation of tissue sections for in situ hybridization.

Performance and quality testing
Endodeoxyribonuclease and exodeoxyribonuclease assays; liquid form tested for absence of RNase activity.

Unit definition
One mAnson unit is described as that amount of enzyme that liberates 1 µmole of Folin-positive amino acid within 1 min at 37°C using hemoglobin as a substrate.

RNAsecure™ RNase Inactivation Reagent Invitrogen™

Ambion® RNAsecure (patents pending) is a unique non-enzymatic reagent that will irreversibly inactivate RNases in solution. 10 mL supplied.
• No post-treatment autoclaving
• Safe, easy-to-use reagent inactivates RNases in solutions
• Inactivation process can be repeated to protect against newly introduced contaminants
• Compatible with downstream procedures, such as RT-PCR and in vitro transcription
• Inactivate RNases in Tris and other solutions that cannot be treated with DEPC RNase contamination in reagents used for RNA isolation and analysis can contribute to experimental inconsistency and, at worst, can cause experimental failure. Traditionally, DEPC has been used to treat solutions that come into contact with RNA. DEPC treatment is time-consuming, possibly hazardous, and only eliminates RNases present at the time of DEPC treatment. In addition, some solutions (e.g., primary amine containing compounds such as Tris) cannot be treated with DEPC at all. Finally, DEPC has to be inactivated by autoclaving post-treatment to prevent it from interfering with downstream enzymatic reactions. RNAsecure Reagent eliminates these problems. Unlike DEPC, RNAsecure can be used on virtually any solution and does not require post-treatment autoclaving. A unique feature of RNAsecure is that reheating after the initial treatment will reactivate the RNase-destroying agent to eliminate any new contaminants. RNAsecure is supplied as a 25X concentrated stock. It is added to solutions as part of reactions (in vitro transcription, RT-PCR, etc.), prior to the addition of enzymes, and heated to 60°C for 10 min to inactivate RNase.

QuantumRNA™ Universal 18S Internal Standard Invitrogen™

The Ambion® QuantumRNA™ 18S Internal Standard with Competimer™ technology uses specially modified Competimer™s of the same sequence as the normal 18S primers that cannot be extended. By adjusting the ratio of 18S Competimer™s to normal 18S mRNA primers, the signal for 18S mRNA can be attenuated to the level of very rare messages. They are used for quantitative end-point RT-PCR and include sufficient reagents for 100 reactions. Because of its invariant expression across tissues and treatments, 18S ribosomal RNA is an ideal internal control for quantitative RNA analysis. The Universal 18S Internal Standards function across the broadest range of eukaryotes, including animals, plants, and many protozoa and produce a 315 bp PCR product.

Accessory Products:
The QuantumRNA™ Classic 18S Internal Standard (SKU# AM1716) produce a different 489 bp PCR product, and the QuantumRNA™ Classic II 18S Internal Standard (SKU# AM1717) produce a 324 bp PCR product.
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