Kit Features The kit is designed for singleplex RT-PCR with SYBR Green dye. For the reverse transcription step, this kit uses a highly efficient Thermophilic Reverse Transcriptase, which is a thermophilic type A polymerase, with optimal temperatures of 60-62°C. The RTase is easily heat-inactivated at ≥90°C for 1min. The RTase efficiently synthesizes a complementary DNA strand on RNA template from a gene-specific primer, ≤1 unit per 20μL of reaction. The RTase reversely-transcribes single digit copies of target RNA molecules consistently. The kit also contains Taq DNA polymerase for PCR with SYBR Green dye. The concentrations of the primers are variable depending on assay designs and thermocycling protocols (Table 1). The preferable PCR product size is ≤150bp. The kit has three formulations of ROX, Low ROX or High ROX concentrations for your choice.  Transportation and Storage The kit can be transported at below 4°C for up to 3 days. The kit should be kept stable in the dark at -20°C for ≤24 months with ≤10 times of freeze-thaw cycles. The kit can be stored at 4°C for a weak. Setup Reaction and Thermocycling Thaw 1-Step 2X RT-PCR Master Mix and other reaction components at room temperature, mix each component, centrifuge and then place on ice. Determine the total volume for the number of reactions, add 5-10% extra volume, and prepare assay mix of all components except RNA template. Mix the assay mix, centrifuge and then place on ice. Aliquot the assay mix into PCR tubes or plate. Add RNA template to PCR tubes or plate. Seal tubes with flat, optically transparent caps or seal plates with optically transparent film. Mix and then briefly centrifuge the tubes or plate. Program PCR instrument with indicated thermo-cycling protocol. Load PCR tubes or plate and start to run. Perform data analysis according to the PCR instrument instructions. Table 1. Setting up a 20μl or 10μl Reaction Component Volume per 20μl Volume per 10μl Final Concentration 2x Mastermix 10μL 5μL 1X Primersa Variable Variable Each 150-900nM RNA Templatesb Variable Variable As low as single digit copies of target RNA to ≤1μg total RNA H2O To 20μL To 10μL Note: Each primer’s Tm should be designed ≥60°C, preferably between 62°C to 65°C, using primer3 software for high efficiency and specificity.   RNA templates should be extracted by a qualified silica-based kit and eluted with low EDTA TE buffer (10mM Tris-HCl, 0.1mM EDTA, pH 8.0-8.3). Table 2. Standard Thermocycling Protocol Stage Temperature Period Number of Cycles I 60°C 10min 1 II 95°C 1min 1 III 95°C 10sec 35-40 III 60°C, signal acquisition 60sec 35-40 IV 60°C to 95°C Various 1 Table 3. Three-Step Thermocycling Protocol Stage Temperature Period Number of Cycles I 60°C 10min 1 II 95°C 1min 1 III 95°C 10sec 35-40 III 60°C 30sec 35-40 III 68-72°C, signal acquisition 30sec 35-40 IV 68°C to 95°C Various 1 Notes: The three-step thermocycling protocol in Table 3 increases overall polymerase activity by 50%, a more effective protocol than Table 2. The primer concentration used in Tables 2 and 3 is typically 0.15-0.2uM. Table 4. Compatible Instruments qPCR Instrument ROX required by instrument Passive dye setup Bio-Rad® iQ™5, CFX96, CFX384, OpticonRoche Lightcycler®Qiagen Rotor-Gene™Eppendorf Mastercycler®Cepheid® SmartCycler® Not recommended Not necessary Applied Biosystems® 7500, 7500 Fast, QuantStudio™, ViiA7™, Agilent Mx™ Low ROX(50nM final concentration) Turn on ROX passive reference dye  Applied Biosystems® 5700, 7000, 7300, 7700, 7900, 7900HT, 7900HT Fast, StepOne™, StepOnePlus™ High ROX(500nM final concentration) Turn on ROX passive reference dye  Precautions If you order a “No ROX” master mix but you have an Applied Biosystems or Thermo Fisher instrument, please turn off ROX passive reference dye button when setup assays.

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2X DNA Loading Buffer is a Ficoll 400-based ready-to-use DNA loading dye for tracking DNA migration during electrophoresis. 2X DNA Loading Buffer contains three dyes with different migration pattern: xylene cyanol FF migrates at approximately 4kb (Blue), bromophenol blue at 300bp (Purple) and orange G at 50bp (Orange) on a 1% agarose gel. Storage: Room temperature or 4 °C

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2X DNA Loading Buffer is a Ficoll 400-based ready-to-use DNA loading dye for tracking DNA migration during electrophoresis. 2X DNA Loading Buffer contains three dyes with different migration pattern: xylene cyanol FF migrates at approximately 4kb (Blue), bromophenol blue at 300bp (Purple) and orange G at 50bp (Orange) on a 1% agarose gel. Storage: Room temperature or 4 °C

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Intended Use The 2x Fast qPCR Mastermix is used for real-time qualitative and quantitative qPCR with SYBR Green dye. ; The Mastermix is a premixed, 2x concentrated solution that has all the components except for gene-specific primers, probes and templates Kit Characterizations The kit is designed for a single gene qPCR with SYBR Green dye. The kit contains Taq-Fast DNA polymerase which can extend more than 300 bases with a short cycling program. The concentrations of the primers and probes are variable depending on specific assays and thermocycling protocols (Table 1). The preferable PCR product size is ≤150bp. The kit has three formulations of No ROX, Low ROX or High ROX concentrations for corresponding instruments(see Table 4). Transportation and storage The kit can be transported at below 4°C for up to 3 days. The kit should be kept stable in the dark at -20°C for ≤24 months with ≤10 times of freeze-thaw cycles. The kit can be stored at 4°C for a week. Setup Reaction and Thermocycling Thaw 2x Mastermix and other reaction components at room temperature, mix each component, centrifuge and then place on ice. Set up reactions (Table 1). Seal tubes with flat, optically transparent caps or seal plate with optically transparent film. Mix and then briefly centrifuge the tubes or plate. Program PCR instrument with indicated thermo-cycling protocol. Load PCR tubes or plate and start to run. Perform data analysis according to the PCR instrument instructions. Table 1. Setting up a 20μl or 10μl Reaction Component Volume per 20μl Volume per 10μl Final Concentration 2x Mastermix 10μL 5μL 1X Primersa Variable Variable Each 150-900nM DNA Templateb Variable Variable ≤30ng genomic DNA/10μL H2O To 20μL To 10μL Note: Each primer’s Tm should be designed ≥60°C, preferably between 62°C to 65°C, using primer3 software for high efficiency and specificity. DNA templates should be extracted by a qualified silica-based kit and eluted with low EDTA TE buffer (10mM Tris-HCl, 0.1mM EDTA, pH 8.0-8.3). Table 2. Standard Thermocycling Protocol Stage Temperature Period Number of Cycles I 95°C 2 min 1 II 95°C 12 sec 35-40 II 60°C, signal acquisition 60 sec 35-40 III 60°C to 95°C  Various 1 Note: The primer concentration used is typically 0.2uM Table 3. Fast Thermocycling Protocol  Stage Temperature Period Number of Cycles I 95°C 1 min 1 II 95°C 5 sec 35-40 II 60°C, signal acquisition 30 sec 35-40 III 60°C to 95°C  Various 1 Note:  The product size for the fast thermocycling protocol is preferred to be less than 90bp. The primer concentration used is typically between 0.4uM and 0.9uM. Table 4. Compatible Instruments qPCR Instrument ROX required by instrument Passive dye setup Bio-Rad® iQ™5, CFX96, CFX384, Opticon Roche Lightcycler®Qiagen Rotor-Gene™Eppendorf Mastercycler®Cepheid® SmartCycler® Not recommended Not necessary Applied Biosystems® 7500, 7500 Fast, QuantStudio™, ViiA7™, Agilent Mx™ Low ROX(50nM final concentration) Turn on ROX passive reference dye  Applied Biosystems® 5700, 7000, 7300, 7700, 7900, 7900HT, 7900HT Fast, StepOne™, StepOnePlus™ High ROX(500nM final concentration) Turn on ROX passive reference dye    Quality Control  Not detectable DNase and RNase contaminations.  Precautions  If you order a “No ROX” master mix but you have an Applied Biosystems or ThermoFisher instrument, please turn off ROX passive reference dye button when setup assays.

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Intended Use The 2x Fast qPCR Mastermix is used for real-time qualitative and quantitative qPCR with SYBR Green dye. ; The Mastermix is a premixed, 2x concentrated solution that has all the components except for gene-specific primers, probes and templates Kit Characterizations The kit is designed for a single gene qPCR with SYBR Green dye. The kit contains Taq-Fast DNA polymerase which can extend more than 300 bases with a short cycling program. The concentrations of the primers and probes are variable depending on specific assays and thermocycling protocols (Table 1). The preferable PCR product size is ≤150bp. The kit has three formulations of No ROX, Low ROX or High ROX concentrations for corresponding instruments(see Table 4). Transportation and storage The kit can be transported at below 4°C for up to 3 days. The kit should be kept stable in the dark at -20°C for ≤24 months with ≤10 times of freeze-thaw cycles. The kit can be stored at 4°C for a week. Setup Reaction and Thermocycling Thaw 2x Mastermix and other reaction components at room temperature, mix each component, centrifuge and then place on ice. Set up reactions (Table 1). Seal tubes with flat, optically transparent caps or seal plate with optically transparent film. Mix and then briefly centrifuge the tubes or plate. Program PCR instrument with indicated thermo-cycling protocol. Load PCR tubes or plate and start to run. Perform data analysis according to the PCR instrument instructions. Table 1. Setting up a 20μl or 10μl Reaction Component Volume per 20μl Volume per 10μl Final Concentration 2x Mastermix 10μL 5μL 1X Primersa Variable Variable Each 150-900nM DNA Templateb Variable Variable ≤30ng genomic DNA/10μL H2O To 20μL To 10μL Note: Each primer’s Tm should be designed ≥60°C, preferably between 62°C to 65°C, using primer3 software for high efficiency and specificity. DNA templates should be extracted by a qualified silica-based kit and eluted with low EDTA TE buffer (10mM Tris-HCl, 0.1mM EDTA, pH 8.0-8.3). Table 2. Standard Thermocycling Protocol Stage Temperature Period Number of Cycles I 95°C 2 min 1 II 95°C 12 sec 35-40 II 60°C, signal acquisition 60 sec 35-40 III 60°C to 95°C  Various 1 Note: The primer concentration used is typically 0.2uM Table 3. Fast Thermocycling Protocol  Stage Temperature Period Number of Cycles I 95°C 1 min 1 II 95°C 5 sec 35-40 II 60°C, signal acquisition 30 sec 35-40 III 60°C to 95°C  Various 1 Note:  The product size for the fast thermocycling protocol is preferred to be less than 90bp. The primer concentration used is typically between 0.4uM and 0.9uM. Table 4. Compatible Instruments qPCR Instrument ROX required by instrument Passive dye setup Bio-Rad® iQ™5, CFX96, CFX384, Opticon Roche Lightcycler®Qiagen Rotor-Gene™Eppendorf Mastercycler®Cepheid® SmartCycler® Not recommended Not necessary Applied Biosystems® 7500, 7500 Fast, QuantStudio™, ViiA7™, Agilent Mx™ Low ROX(50nM final concentration) Turn on ROX passive reference dye  Applied Biosystems® 5700, 7000, 7300, 7700, 7900, 7900HT, 7900HT Fast, StepOne™, StepOnePlus™ High ROX(500nM final concentration) Turn on ROX passive reference dye    Quality Control  Not detectable DNase and RNase contaminations.  Precautions  If you order a “No ROX” master mix but you have an Applied Biosystems or ThermoFisher instrument, please turn off ROX passive reference dye button when setup assays.

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Intended Use The 2x Fast qPCR Mastermix is used for real-time qualitative and quantitative qPCR with SYBR Green dye. ; The Mastermix is a premixed, 2x concentrated solution that has all the components except for gene-specific primers, probes and templates Kit Characterizations The kit is designed for a single gene qPCR with SYBR Green dye. The kit contains Taq-Fast DNA polymerase which can extend more than 300 bases with a short cycling program. The concentrations of the primers and probes are variable depending on specific assays and thermocycling protocols (Table 1). The preferable PCR product size is ≤150bp. The kit has three formulations of No ROX, Low ROX or High ROX concentrations for corresponding instruments(see Table 4). Transportation and storage The kit can be transported at below 4°C for up to 3 days. The kit should be kept stable in the dark at -20°C for ≤24 months with ≤10 times of freeze-thaw cycles. The kit can be stored at 4°C for a week. Setup Reaction and Thermocycling Thaw 2x Mastermix and other reaction components at room temperature, mix each component, centrifuge and then place on ice. Set up reactions (Table 1). Seal tubes with flat, optically transparent caps or seal plate with optically transparent film. Mix and then briefly centrifuge the tubes or plate. Program PCR instrument with indicated thermo-cycling protocol. Load PCR tubes or plate and start to run. Perform data analysis according to the PCR instrument instructions. Table 1. Setting up a 20μl or 10μl Reaction Component Volume per 20μl Volume per 10μl Final Concentration 2x Mastermix 10μL 5μL 1X Primersa Variable Variable Each 150-900nM DNA Templateb Variable Variable ≤30ng genomic DNA/10μL H2O To 20μL To 10μL Note: Each primer’s Tm should be designed ≥60°C, preferably between 62°C to 65°C, using primer3 software for high efficiency and specificity. DNA templates should be extracted by a qualified silica-based kit and eluted with low EDTA TE buffer (10mM Tris-HCl, 0.1mM EDTA, pH 8.0-8.3). Table 2. Standard Thermocycling Protocol Stage Temperature Period Number of Cycles I 95°C 2 min 1 II 95°C 12 sec 35-40 II 60°C, signal acquisition 60 sec 35-40 III 60°C to 95°C  Various 1 Note: The primer concentration used is typically 0.2uM Table 2. Standard Thermocycling Protocol Stage Temperature Period Number of Cycles I 95°C 1 min 1 II 95°C 5 sec 35-40 II 60°C, signal acquisition 30 sec 35-40 III 60°C to 95°C  Various 1 Note:  The product size for the fast thermocycling protocol is preferred to be less than 90bp. The primer concentration used is typically between 0.4uM and 0.9uM. Table 4. Compatible Instruments qPCR Instrument ROX required by instrument Passive dye setup Bio-Rad® iQ™5, CFX96, CFX384, Opticon Roche Lightcycler®Qiagen Rotor-Gene™Eppendorf Mastercycler®Cepheid® SmartCycler® Not recommended Not necessary Applied Biosystems® 7500, 7500 Fast, QuantStudio™, ViiA7™, Agilent Mx™ Low ROX(50nM final concentration) Turn on ROX passive reference dye  Applied Biosystems® 5700, 7000, 7300, 7700, 7900, 7900HT, 7900HT Fast, StepOne™, StepOnePlus™ High ROX(500nM final concentration) Turn on ROX passive reference dye    Quality Control  Not detectable DNase and RNase contaminations.  Precautions  If you order a “No ROX” master mix but you have an Applied Biosystems or ThermoFisher instrument, please turn off ROX passive reference dye button when setup assays.

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Description HiFi PCR Mix for NGS is a premixed system consisting of hot-start enzyme, PCR Buffer, dNTPs, Mg2+, and PCR stabilizers and enhancers. It has high fidelity, high extensibility, and low preference. For complex DNA templates (such as high GC content template), it has a balanced amplification efficiency. This product is particularly suitable for the amplification of multiplex PCR during the construction of NGS library. The highly efficient hot-start enzyme contained in this product does not have polymerase activity at room temperature, thus effectively avoiding non-specific amplification. The combination of a unique buffer system and a hot-start enzyme significantly improves the efficiency of the PCR, resulting in a wider range of amplification. This product effectively increases the amplification efficiency of high GC or high AT regions in the genome, reduces the amplification preference and increases the coverage area of sequencing. Precautions This product cannot use with primers with modifications. Invert gently to mix well before use. Avoid foaming. Use after brief centrifuge. Avoid freezing and thawing repeatedly. Repeated freezing and thawing will comprise the performance of this product. Store at 2-8°C if used frequently.

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An optimized ready-to-use solution containing Hot Start Taq DNA polymerase, standard Hot Start Taq reaction buffer, dNTPs, tracking dye and stabilizers. It is ideally suited to routine PCR applications such as sub-cloning, colony screening and genotyping. It can amplify up to 6 kb from complex genomic DNA. Blue dye migrates approximately at 4 kb. 10x1 ml. Storage: -20ºC ***** This is a cold item. You are requires to select Overnight shipping for purchase this item! *****

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Ready to use solutions optimized for Probe real time PCR (Taqman, Molecular Beacon etc) Include the HotStar DNA Polymerase, dNTPs, ROX and the stabilizing agent and an optimized PCR buffer, ensuring PCR specifity and sensitivity. The ROX in the mix is used for normalization of the flurescent signal in the qPCR. Compatible with real time thermal cyclers for probe real time PCR.

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Ready to use solutions optimized for Probe real time PCR (Taqman, Molecular Beacon etc) Include the HotStar DNA Polymerase, dNTPs, ROX and the stabilizing agent and an optimized PCR buffer, ensuring PCR specifity and sensitivity. The ROX in the mix is used for normalization of the flurescent signal in the qPCR. Compatible with real time thermal cyclers for probe real time PCR.

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Ready to use solutions optimized for Probe real time PCR (Taqman, Molecular Beacon etc) Include the HotStar DNA Polymerase, dNTPs, ROX and the stabilizing agent and an optimized PCR buffer, ensuring PCR specifity and sensitivity. The ROX in the mix is used for normalization of the flurescent signal in the qPCR. Compatible with real time thermal cyclers for probe real time PCR.

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2x Taq PCR Premix is an optimized ready-to-use solution containing Taq DNA polymerase, standard Taq reaction buffer, dNTPs, tracking dyes and stabilizers. It is ideally suited to routine PCR applications such as sub-cloning, colony screening and genotyping. It can amplify up to 4 kb from complex genomic DNA or up to 5 kb from lambda DNA. Tracking dyes included in the mix: Blue dye migrates approximately 4 kb, and yellow dye migrates approximately 50 bp. ***** This is a cold item. You are requires to select Overnight shipping for purchase this item! *****

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