1. RT-PCR Enzymes:

1.1     Thermophilic Reverse Transcriptase (Cat: RT-PCR 140)

Intended use

This reverse transcriptase or RNA-dependent DNA polymerase is used for real-time fluorescent RT-PCR with either SYBR Green dye or TaqMan probe.

Characteristics

• The reverse transcriptase originates from a type A DNA polymerase of thermophilic bacterium (US patent pending).

• This RTase works best at 60-62 °C.

• It is easily heat-inactivated at ≥90 °C for 1 min.

• The enzyme efficiently synthesizes a complementary DNA on RNA template, ≤1 unit per 20 μL reaction.

• The RTase reverse-transcribes single digit copies of target RNA molecules consistently.

• The preferable PCR product size is ≤150 bp.

• It is particularly suitable to one-step real-time qualitative and quantitative RT-PCR.

1.2.     Taq-Probe Polymerase (Cat: RT-PCR 145)

Intended use

• This DNA-dependent DNA polymerase is specially engineered for real-time PCR and RT-PCR with TaqMan probe, singleplex or multiplex.

• It is used together with Thermophilic Reverse Transcriptase (Cat 140) in RT-PCR.

Specification

• Taq-Probe polymerase is specially engineered for TaqMan probe. Its increases 5’-3’ exonuclease activity generating S-shaped fluorescence curve.

• The 5x buffer has three formulations of ROX, Low ROX or High ROX concentrations for your choice.

1.3.     Taq-Fast Polymerase (Cat: RT-PCR 148)

Intended use

• This DNA-dependent DNA polymerase is used real-time PCR and RT-PCR with SYBR Green dye.

• It is used together with Thermophilic Reverse Transcriptase (Cat 140) for RT-PCR.

Characteristics

• Taq-Fast polymerase extends more than 300 bases with short PCR cycling program.

• The 5x buffer has three formulations of ROX, Low ROX or High ROX concentrations for your choice.

Note: Depending on DNA polymerases used in RT-PCR, the kit contains different buffers. Please inquire for detail: contact@genetictoolsllc.com.

a: Taq-Fast polymerase, Cat RT-PCR 148,  extends more than 300 bases with short PCR cycling program.

b: Taq-Probe polymerase, Cat RT-PCR 145, is specially engineered for TaqMan probe, generating S-shaped fluorescence curve.

2. RT-PCR Kits:

2.1.     Extraction-Free 1-Step 2X RT-PCR Master Mix-SYBR Green Dye (Cat: RT-PCR 142)

Intended Use

• This Master Mix is used for real-time qualitative and quantitative RT-PCR from extraction-free RNA samples of tissue-cultured cells.

• The Master Mix is a premixed, 2x concentrated solution that has all the components except for gene-specific primers and RNA template.

Specification

• Using Thermophilic Reverse Transcriptase and Taq-Fast DNA polymerase, this kit contains extra redundant enzyme amounts, optimized to overcome endogenous inhibition in extraction-free RNA samples.

• For the reverse transcription step, this kit uses a highly efficient Thermophilic Reverse Transcriptase (US patent pending), which originates from a type A DNA polymerase of thermophilic bacterium.

• This RTase works best at 60-62 °C.

• It is easily heat-inactivated at ≥90 °C for 1 min.

• The enzyme efficiently synthesizes a complementary DNA on RNA template, 1 unit per 20 μL reaction.

• The RTase reverse-transcribes single digit copies of target RNA molecules consistently.

• The kit also contains Taq-Fast DNA polymerase which extends more than 300 bases with short cycling program.

• The preferable PCR product size is ≤150 bp.

• The kit has three formulations of No ROX, Low ROX or High ROX concentrations for your choice.

2.2. 1-Step 2X RT-PCR Master Mix-TaqMan Probe (Cat: RT-PCR 143)

Intended Use

• The 1-Step 2X RT-PCR Master Mix is used for real-time qualitative and quantitative RT-PCR amplifications with TaqMan probe.

• The Master Mix is a premixed, 2x concentrated solution that has all the components except for gene-specific primers, probe and RNA template.

Specification

• Using Thermophilic Reverse Transcriptase and Taq-Probe DNA polymerase, the kit is designed for singleplex RT-PCR with TaqMan probe.

• For the reverse transcription step, this kit uses a highly efficient Thermophilic Reverse Transcriptase (US patent pending), which originates from a thermophilic type A DNA polymerase.

• This RTase works best at 60-62 °C.

• It is is easily heat-inactivated at ≥90 °C for 1 min.

• The enzyme efficiently synthesizes a complementary DNA on RNA template, ≤1 unit per 20 μL reaction.

• The RTase reverse-transcribes single digit copies of target RNA molecules consistently.

• The kit also contains Taq-Probe DNA polymerase specially engineered for TaqMan probe generating S-shaped fluorescence curve.

• The preferable PCR product size is ≤150 bp.

2.3. 1-Step 2X RT-PCR Master Mix-SYBR Green (Cat: RT-PCR 144)

Intended use

• The 1-Step 2X Fast RT-PCR Master Mix is used for real-time qualitative and quantitative RT-PCR amplifications with SYBR Green dye particularly for fast thermocycling.

• The Master Mix is a premixed, 2x concentrated solution that has all the components except for gene-specific primers and RNA template.

Specification

• Using Thermophilic Reverse Transcriptase and  standard Taq DNA polymerase, 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 (US patent pending), which originates from a thermophilic type A DNA polymerase.

• This RTase works best at 60-62 °C.

• It is easily heat-inactivated at ≥90 °C for 1 min.

• The enzyme efficiently synthesizes a complementary DNA on RNA template, ≤1 unit per 20 μL reaction.

• The RTase reverse-transcribes single digit copies of target RNA molecules consistently.

• The kit also contains standard Taq DNA polymerase for PCR with SYBR Green dye.

• The preferable PCR product size is ≤150 bp.

2.4.     1-Step 2X Multiplex RT-PCR Master Mix-TaqMan Probe (Cat: RT-PCR 146)

Intended Use

• The 1-Step 2X Multiplex RT-PCR Master Mix is used for real-time qualitative and quantitative multiplex RT-PCR amplifications with up to four TaqMan probes.

• The Master Mix is a premixed, 2x concentrated solution that has all the components except for gene-specific primers, probes and RNA templates.

Specification

• Using Thermophilic Reverse Transcriptase and  Taq-Probe DNA polymerase, the kit is designed for multiplex RT-PCR with TaqMan probes.

• For the reverse transcription step, this kit uses a highly efficient Thermophilic Reverse Transcriptase (US patent pending), which originates from a thermophilic type A DNA polymerase.

• This RTase works best 60-62 °C.

• It is easily heat-inactivated at ≥90 °C for 1 min.

• The enzyme efficiently synthesizes complementary DNA on RNA template, 1 unit per 20 μL reaction.

• The RTase reverse-transcribes single digit copies of target RNA molecules consistently.

• The kit also contains Taq-Probe DNA polymerase, specially engineered for TaqMan probes generating S-shaped fluorescence curves.

• Up to four pairs of gene-specific primers can be applied in one reaction.

• The preferable PCR product size is ≤150 bp.

2.5.     1-Step 2X Fast RT-PCR Master Mix-SYBR Green (Cat: RT-PCR 147)

Intended Use

• The 1-Step Fast 2X RT-PCR Master Mix is used for real-time qualitative and quantitative RT-PCR amplifications with SYBR Green dye particularly for fast thermocycling.

• The Master Mix is a premixed, 2x concentrated solution that has all the components except for gene-specific primers and RNA template.

Specification

• Using Thermophilic Reverse Transcriptase and  Taq-Fast DNA polymerase, 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 (US patent pending), which originates from a thermophilic type A DNA polymerase.

• This RTase works best at 60-62 °C.

• It is easily heat-inactivated at ≥90 °C for 1 min.

• The enzyme efficiently synthesizes a complementary DNA on RNA template, ≤1 unit per 20 μL reaction.

• The RTase reverse-transcribes single digit copies of target RNA molecules consistently.

• The kit also contains Taq-Fast DNA polymerase which extends more than 300 bases with short cycling program.

• The preferable PCR product size is ≤150 bp.

2.6. 1-Step 2X Super Multiplex RT-PCR Master Mix-TaqMan Probe (Cat. RT-PCR 149)

Intended Use

• The 1-Step 2X Super Multiplex RT-PCR Master Mix is used for real-time qualitative and quantitative multiplex RT-PCR amplifications with TaqMan probes for up to four targets.

• The Master Mix is a premixed, 2x concentrated solution that has all the components except for gene-specific primers, probes and RNA templates.

Specification

• Using Thermophilic Reverse Transcriptase and  super Taq-Probe DNA polymerase II, the kit is designed for multiplex RT-PCR with TaqMan probes.

• For the reverse transcription step, this kit uses a highly efficient Thermophilic Reverse Transcriptase (US patent pending), which originates from a thermophilic type A DNA polymerase.

• This RTase works best at 60-62 °C.

• It is easily heat-inactivated at ≥90 °C for 1 min.

• The enzyme efficiently synthesizes complementary DNA on RNA template, 1 unit per 20 μL reaction.

• The RTase reverse-transcribes single digit copies of target RNA molecules consistently.

• The kit also contains super Taq-Probe DNA polymerase, specially engineered for TaqMan probes generating S-shaped fluorescence curves.

• Up to four pairs of gene-specific primers can be applied in one reaction.

• The preferable PCR product size is ≤150 bp.

3. qPCR Kits:

3.1.     2X qPCR Master Mix-TaqMan Probe (Cat: PCR 153)

Intended Use

• The 2X qPCR Master Mix is used for real-time qualitative and quantitative qPCR with TaqMan probe.

• The Master Mix is a premixed, 2x concentrated solution that has all the components except for gene-specific primers, probe and DNA template.

Specification

• The kit is designed for singleplex qPCR with TaqMan probe.

• This kit uses Taq-Probe DNA polymerase specially engineered for TaqMan probe, which increases 5’ to 3’ exonuclease efficiency producing S-shaped fluorescence curve.

• The preferable PCR product size is ≤150 bp.

• The kit has three formulations of ROX, Low ROX or High ROX concentrations for your choice (see User Manual Table 2).

3.2.     2X Multiplex qPCR Master Mix-TaqMan Probe (Cat: qPCR 156)

Intended Use

• The 2X Multiplex qPCR Master Mix is used for real-time qualitative and quantitative multiplex qPCR with up to four TaqMan probes.

• The Master Mix is a premixed, 2x concentrated solution that has all the components except for gene-specific primers, probes and  templates.

Specification

• The kit is designed for multiplex qPCR with TaqMan probes.

• The kit uses Taq-Probe DNA polymerase engineered for TaqMan probes, which increases 5’ to 3’ exonuclease efficiency producing S-shaped fluorescence curve.

• Two to four pairs of gene-specific primers can be applied in one reaction.

• The preferable PCR product size is ≤150 bp.

3.3.     2X Fast qPCR Master Mix-SYBR Green (Cat: qPCR 157)

Intended Use

• The 2X Fast qPCR Master Mix is used for real-time qualitative and quantitative qPCR with SYBR Green dye.

• The Master Mix is a premixed, 2x concentrated solution that has all the components except for gene-specific primers and DNA template.

Specification

• The kit is designed for singleplex 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 preferable PCR product size is ≤150 bp.

3.4. 2X Super Multiplex qPCR Master Mix-TaqMan Probe (Cat. qPCR 159)

Intended Use

• The 2X Multiplex Super qPCR Master Mix is used for real-time qualitative and quantitative multiplex qPCR with up to four TaqMan probes.

• The Master Mix is a premixed, 2x concentrated solution that has all the components except for gene-specific primers, probes and  templates.

Specification

• The kit is designed for multiplex qPCR with TaqMan probes.

• The kit uses Super Taq-Probe DNA polymerase specially engineered for TaqMan probes, which increases 5’ to 3’ exonuclease efficiency and  produces S-shaped fluorescence curve.

• Two to four pairs of gene-specific primers can be applied in one reaction.

• The preferable PCR product size is ≤150 bp.

4. PAP Enzymes:

4.1.    PAP Polymerase GP (General Purpose) (Cat: PAP 120)

Intended use

        This DNA-dependent DNA polymerase is used for real-time fluorescent Pyrophosphorolysis Activated Polymerization (PAP) with SYBR Green dye.

Specification

• This polymerase efficiently catalyzes pyrophosphorolysis on DNA template to remove a 3’ dideoxynucleotide or 3’ acyclonucleotide from a blocked primer. 

• It also efficiently synthesizes a DNA on DNA template.

• It detects a single copy of target DNA molecule specifically in PAP.

4.2.    PAP Polymerase HY (High Yield) (Cat: PAP 121)

Intended use

        This DNA-dependent DNA polymerase is used for real-time fluorescent Pyrophosphorolysis Activated Polymerization (PAP) amplification with SYBR Green dye.

Specification

• This polymerase efficiently catalyzes pyrophosphorolysis on DNA template to remove a 3’ dideoxynucleotide or 3’ acyclonucleotide from a blocked primer.

• It also efficiently synthesizes a DNA on DNA template.

• It detects a single copy of target DNA molecule specifically in PAP.

4.3.    PAP Polymerase HP (High Processibility) (Cat: PAP 122)

Intended use

        These DNA-dependent DNA polymerases are used for real-time fluorescent Pyrophosphorolysis Activated Polymerization (PAP) amplification with SYBR Green dye.

Specification

• This polymerase efficiently catalyses pyrophosphorolysis on DNA template to remove a 3’ dideoxynucleotide or 3’ acyclonucleotide from a blocked primer. 

• It also efficiently synthesizes a DNA on DNA template.

• It detects a single copy of target DNA molecule specifically in PAP.

4.4.    RT-PAP Thermophilic Reverse Transcriptase (Cat: PAP 123)

Intended use

        This reverse transcriptase or RNA-dependent DNA polymerase is used for real-time fluorescent RT-Pyrophosphorolysis Activated Polymerization (RT-PAP) with either SYBR Green dye or TaqMan probe.

Specification

• The reverse transcriptase originates from a thermophilic type A polymerase (US patent pending), which originates from a thermophilic type A DNA polymerase.

• This RTase works best at 60-62 °C.

• It is heat-inactivated at ≥90°C for 1 min.

• The RTase efficiently catalyzes pyrophosphorolysis on RNA template to remove a 3’ dideoxynucleotide from a blocked primer. 

• It also efficiently synthesizes a complementary DNA on RNA template, ≤1 unit per 20 μL reaction.

• It detects single digit copies of target RNA molecules specifically in RT-PAP.

4.5.    PAP Taq-Probe Polymerase (Cat: P124)

Intended use

        This DNA-dependent DNA polymerase is used for real-time fluorescent pyrophosphorolysis activated polymerization (PAP) with either singleplex or multiplex TaqMan probes.

Specification

• Particularly, this enzyme is engineered for singleplex or multiplex TaqMan probes.

• This polymerase efficiently catalyses pyrophosphorolysis on DNA template to remove a 3’ dideoxynucleotide or 3’ acyclonucleotide from a blocked primer.

• It also efficiently synthesizes a DNA on DNA template.

• It detects a single copy of target DNA molecule specifically in PAP.

Note: Depending on used DNA polymerases in the amplification step, the kit contains different buffers.  Please Choose a 5x RT-PAP Buffer when you order:

a: PAP polymerase: Cat P120 (GP), P121 (HY) or P122 (HP).

b: PAP Taq-Probe Polymerase: Cat P124, specially engineered for TaqMan probe and generating S-shaped curve in PAP.

5. NA Extraction

5.1. Human Plasma Cell-Free DNA Extraction Kit (Midi 5mL) (Cat: DLC 302)

【Intended use】

• The kit uses a double-layer column to extract circulating cell-free DNA from 5 mL plasma.

• The eluted nucleic acid can be directly used for downstream applications, such as NGS and PCR.

【Specification】

• Sample volume: 5 ml  plasma.

• cfDNA purity: Directly used for downstream applications.

• cfDNA size: 80 to 170 bp. 

• cfDNA recovery rate: ≥90 %.

• Capability: 20 μg.

• No genomic DNA contamination: No gDNA contamination from lysed white cells because it remains bound tightly on the anion exchange membrane without eluted.

• Convenience: No Proteinase K needed.

5.2. Human Plasma Cell-Free DNA Extraction Kit (Mini 0.7mL) (Cat: DLC 303)

【Intended use】

• The kit uses a double-layer column to extract circulating cell-free DNA from 0.7ml of plasma.

• The eluted nucleic acid can be directly used for downstream applications, such as NGS and PCR.

• Swift procedure done in 5 min.

【Specification】

• Sample volume: 0.7ml plasma.

• cfDNA purity: Directly used for downstream applications.

• cfDNA size: In the range of 80 to 170 bp. 

• cfDNA recovery rate: ≥90 %.

• Capability: 20 μg.

• No genomic DNA contamination: No gDNA contamination from lysed white cells because it remains bound tightly on the anion exchange membrane without eluted.

• Convenience: No Proteinase K needed.