Log In Sign up
The largest database of trusted experimental protocols

Superscript 3 taq polymerase

Manufactured by Thermo Fisher Scientific
Visit Supplier
Sourced in United States, United Kingdom

SuperScript III Taq polymerase is a thermostable DNA polymerase enzyme used for DNA amplification in the polymerase chain reaction (PCR) process. It possesses 5' to 3' DNA polymerase activity and 3' to 5' exonuclease activity.

Automatically generated - may contain errors

Lab products found in correlation

Trizol reagent, by Favorgen Biotech (2 mentions) Thermocycler, by Bio-Rad (1 mentions) Ssofast sybr green master mix, by Bio-Rad (1 mentions) Pcr cycler, by Eppendorf (1 mentions) Fluidigm 96.96 dynamicarray ifc, by Standard BioTools (1 mentions) Exonuclease 1, by New England Biolabs (1 mentions) Biomark hd system, by Standard BioTools (1 mentions) Prism 8, by GraphPad (1 mentions) Thermo cycler cfx96, by Bio-Rad (1 mentions)

4 protocols using superscript 3 taq polymerase

1

Real-Time PCR for Influenza Virus Quantification

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Virus titers were quantified by real-time PCR methods as previously reported19 (link),20 (link). Total RNA was isolated from the lungs and BALF using the TRIzol reagent (Favorgen, Ping-Tung, Taiwan). The PCR reaction contained 10 μL of template RNA, standard, or negative control; 12.5 μL of 2 × SuperScript III Platinum Master Mix (Invitrogen); 0.5 μL of SuperScript III Taq polymerase (Invitrogen); and 2 μL each of forward primer (10 μM), reverse primer (10 μM), and dual-labeled probe (5 pmol) in a total volume of 25 μL. Real-time PCR was performed using a Bio-Rad thermocycler (Bio-Rad Laboratories Inc., Hercules, CA, USA). The PCR conditions were as follows: 30 min at 50 °C and 5 min at 95 °C, followed by 45 cycles of 20 s at 95 °C and 1 min at 55 °C. For virus detection, we used two pairs of influenza virus-specific primers and TaqMan probes designed based on the conserved matrix gene region of influenza A virus. The sequences included: forward primer 5′-GACCRATCCTGTCACCTCTGAC-3′, reverse primer 5′-AGGGACTTYTGGACAAAKCGTCTA-3′, and probe 5′-FAM-TGCAGTCCTCGCTCACTGGGCACG-BHQ1-3′.
Bang Y.J., Hong S.H., Park H.J., Kwak H.W., Lee Y.S., Kim J.Y., Park H.J., Bae S.H., Kim H.J., Kim Y.H., Ko H.L., Park S.I., Kim H., Park G., Park M.S., Chang J, & Nam J.H. (2021). Effective inactivated influenza vaccine for the elderly using a single-stranded RNA-based adjuvant. Scientific Reports, 11, 11981.
+ Open protocol
+ Expand
2

Single-cell gene expression profiling

Check if the same lab product or an alternative is used in the 5 most similar protocols
For gene expression analyses, pools of 100 cells were sorted into wells of a DNase and RNase-free 96-well plate (Applied Biosystems) containing 5 μl CellsDirect 2x reaction buffer (Invitrogen), centrifuged for 5 minutes at 500×g, snap-frozen on dry ice and stored at −80°C until use. RNA was reverse-transcribed using Superscript III Taq polymerase (Invitrogen) and preamplified for 18 rounds with a custom 96-target DeltaGene (Fluidigm) primer panel on a PCR cycler (Eppendorf). Excess primers were removed from the preamplified product by incubating with Exonuclease-1 (New England Biolabs), and cDNA samples were diluted in DNA buffer. Primers and cDNAs mixed with SsoFast Sybr Green Master Mix (BioRad) were subsequently loaded onto a Fluidigm 96.96 Dynamic Array IFC and run on a BioMark HD system (Fluidigm). Data were subsequently analyzed using Fluidigm Gene Expression Software and normalized to Gusb. Relative changes were subsequently calculated using ΔΔCt approach. Unsupervised clustering of Gusb-normalized delta CT values with Gusb removed along with poorly performing Ebf1 and Hoxa2 primer sets, was performed using average linkage. Clustering and principal component analysis (PCA), and heatmap generation, were performed using ClustVis software (biit.cs.ut.ee/clustvis). PCA and PCA loading plots were generated using Prism 8 (Graphpad) from data generated by ClustVis.
Rabe J.L., Hernandez G., Chavez J.S., Mills T.S., Nerlov C, & Pietras E.M. (2019). CD34 and EPCR coordinately enrich functional murine hematopoietic stem cells under normal and inflammatory conditions. Experimental hematology.
+ Open protocol
+ Expand
3

Quantitative RT-PCR for Influenza A Virus Detection

Check if the same lab product or an alternative is used in the 5 most similar protocols
Mouse lungs were lavaged using a 22-gauge catheter and 1 mL saline by flushing the airway compartment three times. The BALF was centrifuged at 20,000× g for 10 min at 4 °C. Total RNA from the lungs and BALF was extracted using TRIzol® reagent (Favorgen, Ping-Tung, Taiwan). The PCR reaction mix (25 μL) comprised 12.5 μL 2X SuperScript III Platinum Master Mix (Invitrogen), 2 μL of the mixture comprising forward primer (10 μM), reverse primer (10 μM), and dual-labeled probe (5 pmol), 0.5 μL SuperScript III Taq polymerase (Invitrogen), and 10 μL template RNA, standard, or negative control. Real-time PCR was performed on a Bio-Rad thermocycler CFX96 (Bio-Rad Laboratories Inc., CA, USA). The PCR conditions were as follows: 30 min at 50 °C and 5 min at 95 °C, followed by 45 cycles of 20 s at 95 °C, and 1 min at 55 °C. For virus detection, we used two pairs of influenza virus-specific primers (forward 5′-GACCRATCCTGTCACCTCTGAC-3′, reverse 5′-AGGGACTTYTGGACAAAKCGTCTA-3′) and TaqMan probes (5′-FAM-TGCAGTCCTCGCTCACTGGGCACG-BHQ1) designed based on the conserved matrix gene region of influenza A virus.
Park H.J., Bang Y.J., Kwon S.P., Kwak W., Park S.I., Roh G., Bae S.H., Kim J.Y., Kwak H.W., Kim Y., Yoo S., Kim D., Keum G., Bang E.K., Hong S.H, & Nam J.H. (2023). Analyzing immune responses to varied mRNA and protein vaccine sequences. NPJ Vaccines, 8, 84.
+ Open protocol
+ Expand
4

One-step RT-qPCR for CPMV Detection

Check if the same lab product or an alternative is used in the 5 most similar protocols
The template encapsidated in the CPMV particles was evaluated using two diagnostic RT-qPCR assays (Reid et al., 2002; Callahan et al., 2002 ) using a one-step RT-qPCR protocol previously described (Reid et al., 2009 (link)). Briefly, 5 μl of 10−2–10−9 dilution RNA samples prepared previously was added to 20 μl reaction mix encompassing 12.5 μl of 2× mix (Invitrogen); 2 μl forward primer (10 μM); 2 μl reverse primer (10 μM); 1.5 μl probe (5 μM); 1.5 μl nuclease free water and 0.5 μl SuperScript III/Taq Polymerase (Invitrogen, UK). One step RT-qPCR was performed using the following protocol: one cycle at 60 °C for 30 min; one cycle at 95 °C for 10 min and 50 cycles at 95 °C for 15 s and 60 °C for 1 min. CT (cycle threshold) values were assigned as previously described (Reid et al., 2002 (link)).
Madi M., Mioulet V., King D.P., Lomonossoff G.P, & Montague N.P. (2015). Development of a non-infectious encapsidated positive control RNA for molecular assays to detect foot-and-mouth disease virus. Journal of Virological Methods, 220, 27-34.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!