25u t4 dna polymerase

Manufactured by New England Biolabs

T4 DNA polymerase is a thermostable DNA-dependent DNA polymerase. It catalyzes the addition of deoxynucleotides to the 3' hydroxyl terminus of a DNA primer annealed to a DNA template.

Automatically generated - may contain errors

Lab products found in correlation

Nucleospin pcr clean up kit, by Macherey-Nagel (2 mentions) Megashortscript t7 kit, by Thermo Fisher Scientific (1 mentions) Nucleic acid labeling kit, by Mirus Bio (1 mentions)

Close spelling variants of this product

T4 DNA polymerase (143 citations) DNA polymerase (79 citations) T4 polymerase (10 citations)

2 protocols using 25u t4 dna polymerase

In Vitro Synthesis and Labeling of Pre-miR-181a-1

Check if the same lab product or an alternative is used in the 5 most similar protocols

Pre-miR-181a-1 DNA template was obtained by two oligos (sequence below) after annealing and elongation at 12°C using 25U T4 DNA polymerase (NEB). The DNA template was purified with NucleoSpin PCR Clean-up kit (Macherey-Nagel). Pre-miR-181a-1 in vitro synthesis was performed using the T7 MEGAshortscript kit (Ambion) from 1 µg purified DNA and following manufacturer's instructions, including template removal by DNase TURBO digestion. An amount of 5 µg of pre-miR-181a-1 was labeled with cy3 using the Nucleic Acid Labeling kit (Mirus) following the manufacturer's instructions.
Pre-miR-181a-1_T7_Fw: 5′-TAATACGACTCACTATAGAACATTCAACGCTGTCGGTGAGTTTGGTATCTAAAGGC-3′
Pre-miR-181a-1_Rv: 5′-TGTACAGTCAACGATCGATGGTTTGCCTTTAGATACCAAACTCACCG-3′

Paramasivam P., Stöter M., Corradi E., Dalla Costa I., Höijer A., Bartesaghi S., Sabirsh A., Lindfors L., Yanez Arteta M., Nordberg P., Andersson S., Baudet M.L., Bickle M, & Zerial M. (2022). Quantitative intracellular retention of delivered RNAs through optimized cell fixation and immunostaining. RNA, 28(3), 433-446.

+ Open protocol

+ Expand

In Vitro Synthesis of Pre-miR-181a-1

Check if the same lab product or an alternative is used in the 5 most similar protocols

Pre-miR-181a-1 DNA template was obtained by two oligos (sequence below) after annealing and elongation at 12°C using 25U T4 DNA polymerase (NEB). The DNA template was purified with NucleoSpin PCR Clean-up kit (Macherey-Nagel). Pre-miR-181a-1 in vitro synthesis was performed using T7
MEGAshortscriptTM kit (Ambion) from 1 µg purified DNA and following manufacturer's instructions, including template removal by DNase TURBO digestion. 5 µg pre-miR-181a-1 were labeled with cy3 using Cy5 mRNA were detectable in living cells as soon as 30min of incubation (Figure 1A), only very low signal was retained in fixed cells, suggesting that the fixation protocol fails to quantitatively retain mRNA in the cytoplasm. Second, we quantified the loss of mRNA upon fixation and permeabilization with detergent.

Paramasivam P., Stöter M., Corradi E., Costa I.D., Höijer A., Bartesaghi S., Sabirsh A., Lindfors L., Arteta M.Y., Nordberg P., Andersson S., Baudet M., Bickle M., & Zerial M. (2021). Quantitative intracellular retention of delivered RNAs through optimized cell fixation and immuno-staining.

+ 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?

Revolutionizing how scientists
search and build protocols!