1 cycle sequencing kit

Manufactured by Thermo Fisher Scientific

Sourced in United States

The 1 Cycle Sequencing Kit is a laboratory equipment product designed for DNA sequencing applications. It provides the necessary reagents and components to perform a single cycle of DNA sequencing. The kit includes the essential materials required for the sequencing process, enabling researchers to analyze genetic information.

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Lab products found in correlation

Bigdye terminator v3, by Thermo Fisher Scientific (3 mentions) Topo ta cloning kit, by Thermo Fisher Scientific (1 mentions) Mmessage mmachine t7 transcription kit, by Thermo Fisher Scientific (1 mentions) Platinum gate talen kit, by Addgene (1 mentions) 3100 genetic analyzer, by Thermo Fisher Scientific (1 mentions) Exosap it, by Thermo Fisher Scientific (1 mentions) Abi 3130 genetic analyzer, by Thermo Fisher Scientific (1 mentions) Access rt pcr system, by Promega (1 mentions) Qiaamp viral rna mini kit, by Qiagen (1 mentions) Abi prism 310 genetic analyzer, by Thermo Fisher Scientific (1 mentions) Isohair, by Nippon Gene (1 mentions)

Spelling variants of this product

Big Dye 3.1 cycle sequencing kits (4 citations) ABIPRISM® BigDyeTM Terminator v3.1 Cycle Sequencing Kits (2 citations) BigDye Terminator v.3.1 Cycle Sequencing chemistry kits (2 citations) BigDye Terminator v3.1 Cycles Sequencing Kit (2 citations)

6 protocols using 1 cycle sequencing kit

Plasmid-based TALE Nuclease Generation

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Plasmids coding TALE nucleases were constructed using the Platinum Gate TALEN Kit (1000000043: Addgene, Cambridge, MA, USA)^{26 (link)}. mRNAs coding the TALE nucleases were synthesised using mMESSAGE mMACHINE T7 Transcription kit (AM1344; Thermo Fisher Scientific) and injected into dechorionated unfertilised eggs with a glass capillary at the concentration of 150 ng/μL each with rhodamine dextran as an injection marker (Fig. S1e).
The mutagenic efficiency of TALEN pairs was examined by amplifying the target regions from the genomic DNA extracted from a mixture of 10 tailbud larvae with primers shown in Supplementary Table 4, cloning the products with TOPO TA Cloning Kit (450641; Thermo Fisher Scientific), performing colony PCR with M13For and M13Rev primers, and analysing the sequence of the gel exudates after agarose gel electrophoresis using BigDye Terminator v3.1 Cycle Sequencing Kit.

Nakazawa S., Shirae-Kurabayashi M, & Sawada H. (2019). The role of metalloproteases in fertilisation in the ascidian Ciona robusta. Scientific Reports, 9, 1009.

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Sequencing of LCE3A Enhancer and Luciferase

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LCE3A enhancer (350 base pair) and recombinant luciferase vectors were amplified and sequenced (Supplementary Table S3) on 3100 Genetic Analyzer (Applied Biosystems) using BigDye Terminator, version 3.1, Cycle Sequencing Kit (Thermo Fisher Scientific, Waltham, MA).

Chandra A., Chandra A., Das S., Mazumder S., Senapati S., Chatterjee G, & Chatterjee R. (2021). Functional Mapping of Genetic Interactions between HLA-Cw6 and LCE3A in Psoriasis. The Journal of investigative dermatology, 141(11).

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Molecular Identification of Bacterial Strain BRA-346

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For molecular identification, genomic DNA of the isolated BRA-346 strain was obtained from pure culture as previously described by Pinto et al. (2020) using phenol:chloroform:isoamyl alcohol-based protocol. The gene 16S rRNA was amplified by polymerase chain reaction (PCR) using primers 27F
(5 -AGAGTTTGATCCTGGCTCAG-3 ) e 1494R (5 -ACGGCTACCTTGTTACGACTT-3 ) (Heuer et al., 1997) (link). The PCR reaction was also performed as previously described by Pinto et al. (2020) . The PCR product was stained using SYBR TM Safe DNA Gel (Life Technologies), analyzed by electrophoresis on agarose gel 1%, and purified with ExoSAP-IT (Affymetrix). Sequencing of amplicons was conducted at the Laboratory of Human Molecular Genetics (LGMH), Department of Genetics of Federal University of Pernambuco (UFPE) using Bigdye TM terminator v3.1 cycle sequencing kit (Thermo Fisher). Sequences forward and reverse were analyzed and edited using Geneious Prime 2020.2 softwarefoot_2 . The obtained sequence was deposited in Genbankfoot_3 under the number MW342808 and compared with sequences from EzBioCloud databasefoot_4 .

Furtado L.C., Bauermeister A., Felício R.d., Ortega R., Pinto F.d.C.L., Machado‐Neto J.A., Trivella D.B.B., Pessoa O.D.L., Wilke D.V., Lopes N.P., Jimenez P.C., & Costa‐Lotufo L.V. (2021). Marine Streptomyces sp. Isolated From the Brazilian Endemic Tunicate Euherdmania sp. Produces Dihydroeponemycin and Analogs With Potent Antiglioma Activity. Frontiers in Marine Science, 8.

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Genetic Polymorphism Screening Protocol

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Peripheral blood from the patients and their biological parents and controls was used for DNA extraction. DNA isolation from white blood cells was performed using the standard phenol/chloroform method [27 (link)].
Genotyping of polymorphisms rs4132601, rs2239633, rs3731217, and rs1800566 was performed by polymerase chain reaction – restriction fragment length polymorphism (PCR-RFLP) assay; primer sequences and restriction enzymes are shown in Table I. Genotype of the polymorphism rs10821936 was detected by Sanger sequencing; for the initial PCR we used external primers (F: 5’-CAGTGAGAGCGAGACTGCAC-3’; R: 5’-TCCTGGCAATGTTTTTCACA-3’), while for the second amplification an internal primer was used (F: 5’-ACACCACCCCACATCTCAAT-3’). The direct sequencing of PCR products was performed using ABI Prism BigDye Terminator v3.1 Cycle sequencing kit (Applied Biosystems, Foster city, USA) using the manufacturer's protocol.
Multiplex PCR was performed to detect the presence or absence of GSTT1 and GSTM1 null genotypes [28 (link)].

Kreile M., Piekuse L., Rots D., Dobele Z., Kovalova Z, & Lace B. (2016). Analysis of possible genetic risk factors contributing to development of childhood acute lymphoblastic leukaemia in the Latvian population. Archives of Medical Science : AMS, 12(3), 479-485.

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Enterovirus VP1 Coding Region Amplification

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For RT-PCR amplification, no passage is conducted and total RNA was extracted directly from 140 µl of virus stocks using a QIAamp viral RNA mini kit (Qiagen, Valencia, CA) according to the manufacturer’s procedure. Reverse transcription-PCR (RT-PCR) was performed using an Access RT-PCR System (Promega). Different primer pairs (486/488 and 040/011 for HEV-A, 008/013 and 187/011 for HEV-B) were used to amplify partial VP1 coding region, and the combination of two sequences yielded the entire VP1 coding region [11] (link)–[13] (link). In order to prevent cross-contamination, an RT-PCR using the RNA extracted from normal RD cells served as a blank control, and a negative control containing all the components of the reaction mixture except for the template was also included.
The products were analyzed by agarose gel electrophoresis, and positive products were purified and sequenced directly with a BigDye Terminator v3.1 Cycle Sequencing Kit (Applied Biosystems, Foster City, CA). Sequences were analyzed by an ABI 3130 genetic analyzer (Applied Biosystems). Molecular typing was performed using online Enterovirus Genotyping Tool version 0.1 [14] (link).

Tao Z., Wang H., Li Y., Liu G., Xu A., Lin X., Song L., Ji F., Wang S., Cui N, & Song Y. (2014). Molecular Epidemiology of Human Enterovirus Associated with Aseptic Meningitis in Shandong Province, China, 2006–2012. PLoS ONE, 9(2), e89766.

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Mitochondrial DNA Sequencing from Hair

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Total DNA was extracted from hair shafts by digestion in extraction buffer using ISOHAIR (Code Number 319-03401; Nippon Gene, Tokyo, Japan). The mtDNA spacer D-loop was amplified by PCR using primers M13RV-L15996 and M13(-21)-H408. The analyzed sequences of the D-loop primers were as follows:
mtDNA L15996, 5′-CTCCACCATTAGCACCCAAAGC-3′; and
mtDNA H408, 5′-CTGTTAAAAGTGCATACCGCCA-3′.
The thermocycling profile consisted of an initial denaturation step at 94°C for 1 minute, followed by 32 cycles of 30 seconds at 94°C, 30 seconds at 56°C, and 75 seconds at 72°C. Purified DNA was sequenced in both directions using an ABI PRISM 310 Genetic Analyzer (Applied Biosystems, Foster City, CA, USA) with a BigDye Terminator v3.1 Cycle Sequencing Kit (Applied Biosystems, Foster City, CA, USA).

Nishimura T, & Watanuki S. (2014). Relationship between mitochondrial haplogroup and seasonal changes of physiological responses to cold. Journal of Physiological Anthropology, 33(1), 27.

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