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The Access RT-PCR System is designed for the reverse transcription (RT) and PCR amplification of a specific target RNA from total RNA or mRNA. This one-tube, two-enzyme system provides sensitive, quick and reproducible analysis of even rare RNAs. The system uses AMV Reverse Transcriptase (AMV RT) from Avian Myeloblastosis Virus for first-strand DNA synthesis and thermostable Tfl DNA polymerase from Thermus flavus for second-strand cDNA synthesis and DNA amplification. The Access RT-PCR System includes an optimized single-buffer system that permits extremely sensitive detection of RNA transcripts without a requirement for buffer additions between the reverse transcription and PCR amplification steps. This simplifies the procedure and reduces the potential for contaminating the samples. In addition, the improved performance of AMV Reverse Transcriptase at elevated temperatures in the AMV/Tfl 5X Reaction Buffer minimizes problems encountered with secondary structures in RNA.

Avian Myeloblastosis Virus Reverse Transcriptase (AMV RT) catalyzes DNA polymerization using template DNA, RNA or RNA:DNA hybrids. It requires a primer (DNA primers are more efficient than RNA primers) as well as Mg(2+) or Mn(2+). The enzyme possesses an intrinsic RNase H activity. Both nonionic detergents and sulfhydryl compounds stabilize the enzyme activity in vitro.

High-quality deoxynucleotide triphosphates (dNTPs) are critical for the success of many key procedures such as cDNA synthesis, sequencing and labeling. Promega dNTPs have greater than 99% triphosphate content and are provided at a concentration of 100mM in water at pH 7.5.

High-quality deoxynucleotide triphosphates (dNTPs) are critical for the success of many key procedures such as cDNA synthesis, sequencing and labeling. Promega dNTPs have greater than 99% triphosphate content and are provided at a concentration of 100mM in water at pH 7.5.

High-quality deoxynucleotide triphosphates (dNTPs) are critical for the success of many key procedures such as cDNA synthesis, sequencing and labeling. Promega dNTPs have greater than 99% triphosphate content and are provided at a concentration of 100mM in water at pH 7.5. dUTP (2'-Deoxyuridine, 5'-Triphosphate) can be used in place of dTTP in PCR and RT-PCR protocols to prevent carryover from previous amplifications. The substitution of dUTP for dTTP in PCR results in uracil-containing PCR products that are suitable for most standard applications. The enzyme uracil-N-glycosylase (UNG, also referred to as UDG) can be added to a PCR premix to excise uracil from any contaminating PCR product, thereby preventing false positives. Each lot of dUTP is function-tested to ensure specific DNA amplification and the absence of nuclease activity.

dNTP Mix is a premixed solution containing sodium salts of dATP, dCTP, dGTP and dTTP, each at 10mM in water at pH 7.5; the total concentration of nucleotides is 40mM. One microliter of the dNTP Mix in a 50ul reaction will give a final dNTP concentration of 200uM for each dNTP.

dNTP Mix is a premixed solution containing sodium salts of dATP, dCTP, dGTP and dTTP, each at 10mM in water at pH 7.5; the total concentration of nucleotides is 40mM. One microliter of the dNTP Mix in a 50ul reaction will give a final dNTP concentration of 200uM for each dNTP.

The second generation of GoTaq products, GoTaq G2 Flexi DNA Polymerase reliably amplifies a wide range of PCR templates and provides high-performing results due to improved manufacturing processes, increased reliability and consistency. GoTaq G2 Flexi DNA Polymerase is supplied with 5X Green GoTaq Flexi Buffer and 5X Colorless GoTaq Flexi Buffer and 25mM MgCl2. The GoTaq G2 Flexi DNA Polymerases are supplied in a proprietary formulation containing 50% glycerol with buffers designed for enhanced amplification. The enzyme is a full-length form of Taq DNA polymerase that exhibits 5'-->3' exonuclease activity. The 5X Green GoTaq Reaction and Flexi Buffers contain two dyes (blue and yellow) that separate during electrophoresis to indicate migration progress. The colorless buffer is used when direct fluorescence or absorbance readings are required without prior purification of the amplified DNA from the PCR.

GoTaq G2 Hot Start Master Mixes are ready-to-use mixes containing GoTaq G2 Hot Start Polymerase, buffer, dNTPs and optimized magnesium—you only need to add primers and template and go! The GoTaq G2 Hot Start Green Master Mix also contains a gel loading dye to facilitate downstream gel analysis. The GoTaq G2 Hot Start Colorless Master Mix contains no gel loading dye and is used when downstream applications require fluorescence or absorbance readings without purification. GoTaq G2 DNA Polymerase is bound to a proprietary antibody that blocks polymerase activity until the initial denaturation step of the amplification reaction, allowing hot-start PCR. Hot-start PCR is advantageous for some amplification targets because it may eliminate or minimize primer-dimer and nonspecific products. GoTaq G2 Hot Start Polymerase exhibits 5' --> 3' exonuclease activity.

For superior convenience and improved yield, sensitivity and specificity, choose GoTaq G2 Hot Start Polymerase, which is bound to a proprietary antibody that blocks activity. Activity is restored during initial denaturation, but is inhibited below 70 degrees C, allowing room-temperature reaction setup. GoTaq G2 Hot Start Polymerase is supplied with 5X Green GoTaq Flexi Buffer, 5X Colorless GoTaq Flexi Buffer and 25mM MgCl2. Hot-start PCR is advantageous for some amplification targets because it may eliminate or minimize primer-dimer and nonspecific products. In some cases, hot-start PCR may improve yields. GoTaq G2 Hot Start Polymerase exhibits 5' --> 3' exonuclease activity.

GoTaq Hot Start Master Mixes are premixed, ready-to-use solutions containing GoTaq Hot Start Polymerase, magnesium, dNTPs and buffer. Set up reactions in less than a minute at room temperature; just add your template, water and primers. GoTaq Hot Start Polymerase contains the high-performance GoTaq DNA Polymerase bound to a proprietary antibody that blocks polymerase activity. The polymerase activity is restored during the initial denaturation step when the amplification reactions are heated at 94-95 degrees C for two minutes, enabling hot-start PCR, where polymerase activity is eliminated or minimized at temperatures below 70 degrees C. GoTaq Hot Start Polymerase exhibits 5' --> 3' exonuclease activity. The master mixes are available in green or colorless formats. The green master mix contains a compound that increases sample density, so samples sink easily into wells of an agarose gel, and two dyes (yellow and blue) that separate to allow easy monitoring during electrophoresis.

GoTaq Hot Start Polymerase contains the high-performance GoTaq DNA Polymerase bound to a proprietary antibody that blocks polymerase activity. The polymerase activity is restored during the initial denaturation step when the amplification reactions are heated at 94-95 degrees C for two minutes.

GoTaq Long PCR Master Mix contains the high-performance GoTaq Hot Start Polymerase in a specially formulated mixture with a proprietary thermostable proofreading polymerase. This optimized enzyme mixture allows efficient amplification of up to 40kb from lambda DNA or 30kb from human genomic DNA. The presence of a proofreading enzyme to repair DNA mismatches and a highly processive polymerase allows the polymerase to continue to elongate the DNA much further, resulting in longer DNA amplification. The optimized formulation of the GoTaq Long PCR Master Mix components enables simple reaction setup and provides consistently efficient, accurate and robust amplification of long DNA amplicons.

The 5X Green GoTaq Reaction Buffer contains two dyes (a blue dye and a yellow dye) that separate during electrophoresis to show migration progress. The buffer also contains a compound that increases sample density. This means that samples can be loaded directly onto gels without the need for loading dye. The blue dye migrates at the same rate as a 3-5kb DNA fragment in a 1% agarose gel. The yellow dye migrates at a rate faster than primers (<50bp) in a 1% agarose gel. The 5X Colorless GoTaq Reaction Buffer has the same formulation as the 5X Green GoTaq Reaction Buffer but does not contain dyes and is recommended for any applications where absorbance or fluorescence measurements are necessary prior to PCR cleanup. Both buffers are supplied at pH 8.5.

The ImProm-II Reverse Transcription System produces efficient, robust synthesis of first-strand cDNA in preparation for PCR amplification. The components of the ImProm-II Reverse Transcription System can be used to reverse transcribe RNA templates starting with total RNA, poly(A)+ mRNA or synthetic transcript RNA. The optimized reaction buffer and powerful ImProm-II Reverse Transcriptase provided in the ImProm-II System together enable robust, full-length cDNA synthesis for the reproducible analysis of rare or long messages. The cDNA synthesis conditions were formulated for standalone applications or for easy transition to gene-specific target amplification. An aliquot of the reverse transcription reaction (1-20ul) can be amplified directly using Taq DNA polymerase in coupled or uncoupled PCR.

Moloney Murine Leukemia Virus Reverse Transcriptase (M-MLV RT) is an RNA-dependent DNA polymerase that can be used in cDNA synthesis with long messenger RNA templates (>5kb). The enzyme is a product of the pol gene of M-MLV and consists of a single subunit with a molecular weight of 71kDa. The RNase H activity of M-MLV RT is weaker than that of the commonly used Avian Myeloblastosis Virus (AMV) reverse transcriptase.

Moloney Murine Leukemia Virus Reverse Transcriptase, RNase H Minus (M-MLV RT [H-]), is an RNA-dependent DNA polymerase that can be used in cDNA synthesis with long messenger RNA templates (>5kb). This form of M-MLV Reverse Transcriptase has been genetically altered to remove the associated RNase H activity. Although many researchers are successful in using M-MLV RT (H+) for analytical and some preparative cDNA applications, reverse transcriptases lacking RNase H activity provide another option to prepare long cDNAs and libraries containing a high percentage of full-length cDNA.

Moloney Murine Leukemia Virus Reverse Transcriptase, RNase H Minus (M-MLV RT [H-]), Point Mutant, is an RNA-dependent DNA polymerase that can be used in cDNA synthesis with long RNA templates (>5kb). The lack of RNase H activity is beneficial for this application, as RNase H can start to degrade templates when incubation times are long, as they may be when synthesizing long cDNAs. Although many researchers are successful in using M-MLV RT (H+) for analytical and some preparative cDNA applications, reverse transcriptases lacking RNase H activity provide another option to prepare long cDNAs and libraries containing a high percentage of full-length cDNA.

The Reverse Transcription System provides reagents to efficiently reverse transcribe RNA into cDNA in 15 minutes. The cDNA prepared from each reaction using this system may be used directly in multiple PCR amplifications using Taq DNA polymerase. The AMV Reverse Transcriptase synthesizes single-stranded cDNA from total or poly(A)+ RNA. Both Oligo(dT)15 and Random Primers are included, allowing cDNA synthesis from virtually any RNA source. The system contains sufficient reagents for 100 cDNA synthesis reactions, processing 1ug of RNA per reaction. Each cDNA synthesis reaction may be divided and used in up to 20 separate PCR amplifications. A polyadenylated 1.2kb RNA transcript is provided as a control template for cDNA synthesis.

Native and Recombinant RNasin Inhibitors are 50kDa proteins that inhibit RNase A family and human placental RNases by noncovalently binding to RNases in a 1:1 ratio. For downstream applications such as GoScript Reverse Transcriptase, AMV/M-MLV reverse transcriptases, SP6, T7/T3 RNA polymerase, and Taq DNA polymerases, Recombinant RNasin Inhibitor does not inhibit RNase T1, S1 nuclease, RNase from Aspergillus, RNase H, RNase ONE Ribonuclease and enzymes. RNasin Plus RNase Inhibitor is a recombinant mammalian RNase inhibitor that is expressed as a soluble protein in E. coli, allowing easy purification through a combination of ion exchange and hydrophobic interaction chromatography. The protein is capable of inhibiting eukaryotic RNases (e.g., RNase A and RNase B) similarly to human placental RNase inhibitor. RNasin Plus RNase Inhibitor is tested in RT-PCR and compatible with enzymes such as AMV, M-MLV and ImProm-II Reverse Transcriptases or Taq and Tfl DNA Polymerases.

High-quality deoxynucleotide triphosphates (dNTPs) are critical for the success of many key procedures such as cDNA synthesis, sequencing and labeling. Promega dNTPs have greater than 99% triphosphate content and are provided at a concentration of 100mM in water at pH 7.5.

High-quality deoxynucleotide triphosphates (dNTPs) are critical for the success of many key procedures such as cDNA synthesis, sequencing and labeling. Promega dNTPs have greater than 99% triphosphate content and are provided at a concentration of 100mM in water at pH 7.5. dUTP (2'-Deoxyuridine, 5'-Triphosphate) can be used in place of dTTP in PCR and RT-PCR protocols to prevent carryover from previous amplifications. The substitution of dUTP for dTTP in PCR results in uracil-containing PCR products that are suitable for most standard applications. The enzyme uracil-N-glycosylase (UNG, also referred to as UDG) can be added to a PCR premix to excise uracil from any contaminating PCR product, thereby preventing false positives. Each lot of dUTP is function-tested to ensure specific DNA amplification and the absence of nuclease activity.

High-quality deoxynucleotide triphosphates (dNTPs) are critical for the success of many key procedures such as cDNA synthesis, sequencing and labeling. Promega dNTPs have greater than 99% triphosphate content and are provided at a concentration of 100mM in water at pH 7.5. dUTP (2'-Deoxyuridine, 5'-Triphosphate) can be used in place of dTTP in PCR and RT-PCR protocols to prevent carryover from previous amplifications. The substitution of dUTP for dTTP in PCR results in uracil-containing PCR products that are suitable for most standard applications. The enzyme uracil-N-glycosylase (UNG, also referred to as UDG) can be added to a PCR premix to excise uracil from any contaminating PCR product, thereby preventing false positives. Each lot of dUTP is function-tested to ensure specific DNA amplification and the absence of nuclease activity.