Log In Sign up
The largest database of trusted experimental protocols

Abi prism bigdye terminator cycle sequencing v 2.0 ready reaction kit

Manufactured by Thermo Fisher Scientific
Visit Supplier
Sourced in United States

The ABI PRISM BigDye Terminator Cycle Sequencing v.2.0 Ready Reaction kit is a reagent kit used for DNA sequencing. The kit includes the necessary components for conducting cycle sequencing reactions, which is a method for determining the nucleotide sequence of DNA molecules.

Automatically generated - may contain errors

Lab products found in correlation

Abi prism 3730 dna analyzer, by Thermo Fisher Scientific (7 mentions) Nucleofast 96 pcr plates, by Takara Bio (1 mentions) Pcr4 topo vector, by Thermo Fisher Scientific (1 mentions) Proteinase k, by Thermo Fisher Scientific (1 mentions) Exosap it, by Thermo Fisher Scientific (1 mentions) Alamut visual version 2, by Sophia Genetics (1 mentions) Superscript vilo master mix, by Thermo Fisher Scientific (1 mentions) Nucleospin rna clean up kit, by Macherey-Nagel (1 mentions)

Spelling variants of this product

ABI PRISM BigDye Terminator Cycle Sequencing Ready Reaction Kit (133 citations) ABI Prism BigDye Terminator Cycle Sequencing kit (100 citations) ABI BigDye Terminator Cycle Sequencing Kit (18 citations) BigDye Terminator Cycle Sequencing Kit (v.2.0; (6 citations) ABI Prism BigDye Terminator cycle sequencing (6 citations) ABI Prism Terminator Cycle Sequencing kit (6 citations) ABI PRISM BigDye Terminator Cycle Sequencing Ready Reaction (3 citations) ABI PRISM BigDye Terminator cycle (3 citations) ABI PRISM BigDye Terminator Cycle Sequencing Ready Kit (2 citations) ABI Prism® BigDye™ cycle sequencing kit (2 citations)

7 protocols using abi prism bigdye terminator cycle sequencing v 2.0 ready reaction kit

1

NGS Variant Validation by Sanger

Check if the same lab product or an alternative is used in the 5 most similar protocols
Mutations detected by NGS were verified by Sanger sequencing using the ABI PRISM BigDye Terminator Cycle Sequencing v.2.0 Ready Reaction kit and ABI PRISM 3730 DNA analyzer (Applied Biosystems). MLPA was used to detect large insertions or deletions in patients with only one or no mutations detected as previously described [46 (link)].
Ivanov M., Matsvay A., Glazova O., Krasovskiy S., Usacheva M., Amelina E., Chernyak A., Ivanov M., Musienko S., Prodanov T., Kovalenko S., Baranova A, & Khafizov K. (2018). Targeted sequencing reveals complex, phenotype-correlated genotypes in cystic fibrosis. BMC Medical Genomics, 11(Suppl 1), 13.
+ Open protocol
+ Expand
2

Sanger Sequencing of CUL4B Gene Variants

Check if the same lab product or an alternative is used in the 5 most similar protocols
Validation of variants and sequencing of the coding exons of the
CUL4B gene was performed using Sanger sequencing on DNA extracted from
peripheral blood. Primer sequences and conditions for PCR are available upon request. PCR
products were sequenced using the ABI PRISM BigDye Terminator Cycle Sequencing V2.0 Ready
Reaction Kit and analyzed with the ABI PRISM 3730 DNA analyzer (Applied Biosystems, Foster
City, CA). DNA of all available family members was analyzed for the variant found in the
index patient to confirm the segregation of the variant with the disease. Nucleotide
numbering uses +1 as the A of the ATG translation initiation codon in the
reference sequence (NM_003588.3, NP_003579.3), with the initiation codon as codon 1. All
variants identified in this study have been submitted to http://www.lovd.nl/CUL4B.
Vulto-van Silfhout A.T., Nakagawa T., Bahi-Buisson N., Haas S.A., Hu H., Bienek M., Vissers L.E., Gilissen C., Tzschach A., Busche A., Müsebeck J., Rump P., Mathijssen I.B., Avela K., Somer M., Doagu F., Philips A.K., Rauch A., Baumer A., Voesenek K., Poirier K., Vigneron J., Amram D., Odent S., Nawara M., Obersztyn E., Lenart J., Charzewska A., Lebrun N., Fischer U., Nillesen W.M., Yntema H.G., Järvelä I., Ropers H.H., de Vries B.B., Brunner H.G., van Bokhoven H., Raymond F.L., Willemsen M.A., Chelly J., Xiong Y., Barkovich A.J., Kalscheuer V.M., Kleefstra T, & de Brouwer A.P. (2015). Variants in CUL4B are Associated with Cerebral Malformations. Human mutation, 36(1), 106-117.
+ Open protocol
+ Expand
3

FoxP Isoforms Amplification and Sequencing

Check if the same lab product or an alternative is used in the 5 most similar protocols
Primers for amplification of FoxP isoforms were designed with Primer3 (http://bioinfo.ut.ee/primer3-0.4.0/). Amplification by PCR was performed on 40 ng of cDNA with Taq DNA polymerase Amplitaq (Life Technologies). Primer sequences are provided in S1 Table. PCR fragments were purified with NucleoFast 96 PCR plates (Clontech) in accordance with the manufacturer's protocol. Sequence analysis was performed with the ABI PRISM BigDye Terminator Cycle Sequencing V2.0 Ready Reaction kit and analyzed with an ABI PRISM 3730 DNA analyzer (Applied Biosystems).
Castells-Nobau A., Eidhof I., Fenckova M., Brenman-Suttner D.B., Scheffer-de Gooyert J.M., Christine S., Schellevis R.L., van der Laan K., Quentin C., van Ninhuijs L., Hofmann F., Ejsmont R., Fisher S.E., Kramer J.M., Sigrist S.J., Simon A.F, & Schenck A. (2019). Conserved regulation of neurodevelopmental processes and behavior by FoxP in Drosophila. PLoS ONE, 14(2), e0211652.
+ Open protocol
+ Expand
4

Sanger Sequencing for Variant Validation

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Variants classified as uncertain by manual curation and pathogenic, likely pathogenic, or VUS based on clinical interpretation were validated by Sanger sequencing. Sanger sequencing was performed using the ABI PRISM BigDye Terminator Cycle Sequencing v.2.0 Ready Reaction kit and ABI PRISM 3730 DNA analyzer (Applied Biosystems) as previously described [26 (link)]. Blood samples were used for Sanger sequencing. All blood samples matched the corresponding tumor samples.
Lebedeva A., Shaykhutdinova Y., Seriak D., Ignatova E., Rozhavskaya E., Vardhan D., Manicka S., Sharova M., Grigoreva T., Baranova A., Mileyko V, & Ivanov M. (2022). Incidental germline findings during molecular profiling of tumor tissues for precision oncology: molecular survey and methodological obstacles. Journal of Translational Medicine, 20, 29.
+ Open protocol
+ Expand
5

Genomic DNA Extraction and ush2a Exon Sequencing

Check if the same lab product or an alternative is used in the 5 most similar protocols
Genomic DNA was isolated from pools of 15 larvae after incubation in 75 μl lysis buffer (10 mM Tris HCl pH=8.2, 10 mM EDTA, 100 mM NaCl and 0.5 % SDS) supplemented with freshly added proteinase K (final concentration of 0.20 mg/ml, Invitrogen #25530049) at 55°C for two hours. The isolated genomic DNA was subsequently used as a template in a PCR. Primers used for the amplification of ush2a exon13 are 5′-TCCACCAACAGAATCTAAATCTTTC-3′ and 5′-CTGATTTGTAAATGGTGTTGGG-3′ and primers used for the amplification of ush2a exon71 are 5′-CATGTTTTGGTTATCTGTTCTTCT-3′ and 5′-GACAGCGGAATGGTGAGATAAAC-3′. The obtained amplicons were cloned into a pCR®4-TOPO® vector (Invitrogen, #450030) according to manufacturer’s instructions. Individual clones were analyzed for the presence of mutational events using the ABI PRISM Big Dye Terminator Cycle Sequencing V2.0 Ready Reaction kit and the ABIPRISM 3730 DNA analyzer (Applied Biosystems).
Dona M., Slijkerman R., Lerner K., Broekman S., Wegner J., Howat T., Peters T., Hetterschijt L., Boon N., de Vrieze E., Sorusch N., Wolfrum U., Kremer H., Neuhauss S., Zang J., Kamermans M., Westerfield M., Phillips J, & van Wijk E. (2018). Usherin defects lead to early-onset retinal dysfunction in zebrafish. Experimental eye research, 173, 148-159.
+ Open protocol
+ Expand
6

Sequence Analysis of Genetic Variants

Check if the same lab product or an alternative is used in the 5 most similar protocols
PCR of fragments to be analyzed was performed using standard protocols. Primers for PCR were designed with Primer3Plus. Reference sequence identifiers, primer sequences and PCR conditions are provided in Table S3. PCR fragments were purified with ExoSAP-IT (Thermo Fisher Scientific), according to manufacturers’ protocols. For sequence analysis, the ABI PRISM BigDye Terminator Cycle Sequencing v2.0 Ready Reaction kit and the ABI PRISM 3730 DNA analyzer (Applied Biosystems, Foster City, CA, USA) were employed. Possibly deleterious effects of the identified variants were predicted with Alamut Visual version 2.7.1 (Interactive Biosoftware, Rouen, France).
Wesdorp M., de Koning Gans P.A., Schraders M., Oostrik J., Huynen M.A., Venselaar H., Beynon A.J., van Gaalen J., Piai V., Voermans N., van Rossum M.M., Hartel B.P., Lelieveld S.H., Wiel L., Verbist B., Rotteveel L.J., van Dooren M.F., Lichtner P., Kunst H.P., Feenstra I., Admiraal R.J., Yntema H.G., Hoefsloot L.H., Pennings R.J, & Kremer H. (2018). Heterozygous missense variants of LMX1A lead to nonsyndromic hearing impairment and vestibular dysfunction. Human Genetics, 137(5), 389-400.
+ Open protocol
+ Expand
7

RNA Isolation and RT-PCR Analysis of MED12 Splicing

Check if the same lab product or an alternative is used in the 5 most similar protocols
All RNA isolations were performed with the NucleoSpin RNA Clean-up Kit (catalog number 740955-50, Macherey-Nagel, Düren, Germany) according to manufacturer's protocol. RNA was quantified by nanodrop. One microgram of total RNA was used for all complementary DNA (cDNA) synthesis reactions. For reverse transcription PCR (RT-PCR) analysis, cDNA was synthesized by the SuperScript VILO Master Mix (catalog number 11755050, Thermo Fisher Scientific, Waltham, MA, USA), following manufacturer's instructions.
To determine the potential splice acceptor variant effect on MED12 splicing in individuals 6 and 7, RT-PCR analysis was performed using primers located in exon 8 (forward) and exon 12 (reverse) of MED12. Primer sequences are available upon request. For RT-PCR analysis, 25 ng of cDNA was used for all reactions. Amplifications were performed according to standard procedures. PCR products were sequenced using the ABI PRISM BigDye Terminator Cycle Sequencing V2.0 Ready Reaction Kit and analyzed with the ABI PRISM 3730 DNA analyzer (Applied Biosystems, Foster City, CA, USA).
Polla D.L., Bhoj E.J., Verheij J.B.G.M., Wassink-Ruiter J.S.K., Reis A., Deshpande C., Gregor A., Hill-Karfe K., Silfhout A.T.V., Pfundt R., Bongers E.M.H.F., Hakonarson H., Berland S., Gradek G., Banka S., Chandler K., Gompertz L., Huffels S.C., Stumpel C.T.R.M., Wennekes R., Stegmann A.P.A., Reardon W., Leenders E.K.S.M., de Vries B.B.A., Li D., Zackai E., Ragge N., Lynch S.A., Cuddapah S., van Bokhoven H., Zweier C, & de Brouwer A.P.M. (2021). De novo variants in MED12 cause X-linked syndromic neurodevelopmental disorders in 18 females. Genetics in medicine : official journal of the American College of Medical Genetics, 23(4).
+ 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!