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Terminal transferase

Manufactured by PerkinElmer
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Terminal transferase is an enzyme that catalyzes the addition of nucleotides to the 3' end of DNA strands. It is commonly used in molecular biology and genetic research applications.

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Lab products found in correlation

Biotin 11 ddatp, by PerkinElmer (1 mentions) α 32p datp, by PerkinElmer (1 mentions) Klenow fragment, by PerkinElmer (1 mentions) T4 polynucleotide kinase, by PerkinElmer (1 mentions) γ 32p atp, by PerkinElmer (1 mentions)

3 protocols using terminal transferase

1

Synthesis of Poly-A Tailed DNA Barcodes

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Thirty types of poly-A tailed DNA barcode molecules were prepared by referring to the sequence of Helicos control oligo. Firstly, we added a poly-A tail to the oligonucleotides (Supplementary Table 1) by incubating 10 µl of mixture consisting of 10 µM oligonucleotide mix, 1× TdT buffer, 250 µM CoCl2, 240 µM dATP, and 1 U/µl of terminal transferase at 37 °C for 1 h and 70 °C for 10 min followed by a 4-°C hold. Secondly, the sample was added to 10 µl of mixture consisting of 1× TdT buffer, 250 µM CoCl2, 160 µM biotin-11-ddATP (PerkinElmer), and 2 U/µl terminal transferase (NEB) and incubated at 37 °C for 1 h, and 70 °C for 10 min followed by a 4-°C hold.
Oguchi Y., Shintaku H, & Uemura S. (2020). Development of a sequencing system for spatial decoding of DNA barcode molecules at single-molecule resolution. Communications Biology, 3, 788.
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2

Radioactive Labeling of CRISPR Substrates

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All oligonucleotides were synthesized by Integrated DNA Technologies. Sequences of all DNA substrates are shown in Table S2. Prespacers and minimal dsDNA CRISPR arrays were hybridized by heating to 95 °C for 5 minutes and slow cooling to room temperature in oligo annealing buffer (20 mM HEPES (pH 7.5), 25 mM KCl, 10 mM MgCl2) and purified on 8% native PAGE. Prespacers were labeled with [γ-32P]-ATP (PerkinElmer) and T4 polynucleotide kinase (NEB) for 5'-end labelling or with [α-32P]-dATP (PerkinElmer) and Terminal Transferase (NEB) for 3'-end labelling. The double-stranded minimal CRISPRs were labeled with [α-32P]-dATP (PerkinElmer) and Klenow-fragment (NEB) for 3'-end labelling.
Lee H., Zhou Y., Taylor D.W, & Sashital D.G. (2018). Cas4-dependent prespacer processing ensures high-fidelity programming of CRISPR arrays. Molecular cell, 70(1), 48-59.e5.
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3

Radiolabeling of DNA Substrates

Cited 1 time
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DNA substrates were 5′ end-labeled with [γ-32P]ATP and T4 polynucleotide kinase as we described (32 (link)). DNA substrates were 3′ 32P-labeled by incubating 40 pmol of the appropriate SS DNA with 30 units of terminal transferase (Perkin Elmer) in the presence of [α-32P]dCTP according to the manufacturer's protocol. The 32P-labeled SS DNA was annealed to non-labeled DNAs to prepare indicated DNA substrates for DNA cleavage assay.
Kim H.S., Nickoloff J.A., Wu Y., Williamson E.A., Sidhu G.S., Reinert B.L., Jaiswal A.S., Srinivasan G., Patel B., Kong K., Burma S., Lee S.H, & Hromas R.A. (2017). Endonuclease EEPD1 Is a Gatekeeper for Repair of Stressed Replication Forks. The Journal of Biological Chemistry, 292(7), 2795-2804.
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