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Gs junior titanium empcr kit lib a kit

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The GS Junior Titanium emPCR kit (Lib-A kit) is a laboratory equipment product designed for use with the GS Junior Sequencing System. The kit is used to perform emulsion PCR (emPCR) amplification of DNA library samples in preparation for sequencing on the GS Junior platform.

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Lab products found in correlation

Gs junior sequencer, by Roche (4 mentions) Thequant it picogreen dsdna reagent, by Thermo Fisher Scientific (2 mentions) Gs amplicon variantanalyzer version 3.0 avav3.0 software, by Roche (2 mentions) Quant it picogreen dsdna reagent, by Thermo Fisher Scientific (2 mentions) Gs junior sequencing, by Roche (2 mentions) Agencourt ampure xp, by Beckman Coulter (2 mentions)

Close spelling variants of this product

GS Junior Titanium emPCR Kit (Lib-A) GS Junior Titanium emPCR Kit ( GS emPCR kit GS Junior Lib-A

4 protocols using gs junior titanium empcr kit lib a kit

1

Screening for PIK3R2 p.Gly373Arg Mutation

Check if the same lab product or an alternative is used in the 5 most similar protocols
To screen for the recurrent PIK3R2 mutation,
p.Gly373Arg, we performed locus-specific amplification of genomic DNA
followed by GS Junior sequencing. We designed fusion primers containing
genome-specific sequences along with distinct MIDs (multiplex identifier
sequences) used to differentiate samples being run together on the same
plate and sequencing adapters to generate amplicons ranging in size from 290
to 310 bp using primer3plus software. Primer sequences are available upon
request (Dr. Renzo Guerrini). Small DNA fragments were removed using
Agencourt AMPure XP (Beckman Coulter, Beverly, MA) according to the
manufacturer’s protocol. All amplicons were quantified using the
Quant-iT PicoGreen dsDNA reagent (Invitrogen Corporation, Life Technologies,
Carlsbad, CA), pooled at equimolar ratios, amplified by emulsion PCR using
the GS Junior Titanium emPCR kit (Lib-A kit, Roche Applied Science,
Mannheim, Germany) and pyrosequenced in the sense and antisense strands on a
GS Junior sequencer (Roche) following the manufacturer’s
instructions. We performed data analysis using the GS Amplicon Variant
Analyzer version 3.0 (AVAv3.0) software (Roche).
Mirzaa G., Conti V., Timms A.E., Smyser C.D., Ahmed S., Carter M., Barnett S., Hufnagel R.B., Goldstein A., Narumi-Kishimoto Y., Olds C., Collins S., Johnston K., Deleuze J.F., Nitschké P., Friend K., Harris C., Goetsch A., Martin B., Boyle E.A., Parrini E., Mei D., Tattini L., Slavotinek A., Blair E., Barnett C., Shendure J., Chelly J., Dobyns W.B, & Guerrini R. (2015). Characterization of mutations of the phosphoinositide-3-kinase regulatory subunit, PIK3R2, in perisylvian polymicrogyria: a next generation sequencing study. The Lancet. Neurology, 14(12), 1182-1195.
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2

Amplicon Sequencing of Genomic DNA

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Amplicon sequencing was performed using previously published methods (48 (link)). We performed locus-specific amplification of genomic DNA followed by GS Junior sequencing (Roche). We designed fusion primers containing genome-specific sequences along with distinct multiplex identifier sequences (used to differentiate samples being run together on the same plate) and sequencing adapters, to generate amplicons ranging in size from 290 to 310 bp, using Primer3Plus software. Primer sequences are provided in Supplemental Table 4. Small DNA fragments were removed with Agencourt AMPure XP (Beckman Coulter) according to the manufacturer’s protocol. All amplicons were quantified with the Quant-iT PicoGreen dsDNA reagent (Life Technologies), pooled at equimolar ratios, amplified by emulsion PCR using the GS Junior Titanium emPCR kit (Lib-A kit, Roche Applied Science), and pyrosequenced in the sense and antisense strands on a GS Junior sequencer following the manufacturers’ instructions. Data were analyzed using GS Amplicon Variant Analyzer version 3.0 software.
Mirzaa G., Timms A.E., Conti V., Boyle E.A., Girisha K.M., Martin B., Kircher M., Olds C., Juusola J., Collins S., Park K., Carter M., Glass I., Krägeloh-Mann I., Chitayat D., Parikh A.S., Bradshaw R., Torti E., Braddock S., Burke L., Ghedia S., Stephan M., Stewart F., Prasad C., Napier M., Saitta S., Straussberg R., Gabbett M., O’Connor B.C., Keegan C.E., Yin L.J., Lai A.H., Martin N., McKinnon M., Addor M.C., Boccuto L., Schwartz C.E., Lanoel A., Conway R.L., Devriendt K., Tatton-Brown K., Pierpont M.E., Painter M., Worgan L., Reggin J., Hennekam R., Tsuchiya K., Pritchard C.C., Aracena M., Gripp K.W., Cordisco M., Van Esch H., Garavelli L., Curry C., Goriely A., Kayserilli H., Shendure J., Graham J J.r., Guerrini R, & Dobyns W.B. (2016). PIK3CA-associated developmental disorders exhibit distinct classes of mutations with variable expression and tissue distribution. JCI Insight, 1(9), e87623.
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3

Amplicon Sequencing for Genomic Analysis

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Amplicon sequencing was performed using previously published methods (48 (link)). We performed locus-specific amplification of genomic DNA followed by GS Junior sequencing (Roche). We designed fusion primers containing genome-specific sequences along with distinct multiplex identifier sequences (used to differentiate samples being run together on the same plate) and sequencing adapters, to generate amplicons ranging in size from 290 to 310 bp, using Primer3Plus software. Primer sequences are provided in Supplemental Table 4. Small DNA fragments were removed with Agencourt AMPure XP (Beckman Coulter) according to the manufacturer’s protocol. All amplicons were quantified with the Quant-iT PicoGreen dsDNA reagent (Life Technologies), pooled at equimolar ratios, amplified by emulsion PCR using the GS Junior Titanium emPCR kit (Lib-A kit, Roche Applied Science), and pyrosequenced in the sense and antisense strands on a GS Junior sequencer following the manufacturers’ instructions. Data were analyzed using GS Amplicon Variant Analyzer version 3.0 software.
Mirzaa G., Timms A.E., Conti V., Boyle E.A., Girisha K.M., Martin B., Kircher M., Olds C., Juusola J., Collins S., Park K., Carter M., Glass I., Krägeloh-Mann I., Chitayat D., Parikh A.S., Bradshaw R., Torti E., Braddock S., Burke L., Ghedia S., Stephan M., Stewart F., Prasad C., Napier M., Saitta S., Straussberg R., Gabbett M., O’Connor B.C., Keegan C.E., Yin L.J., Lai A.H., Martin N., McKinnon M., Addor M.C., Boccuto L., Schwartz C.E., Lanoel A., Conway R.L., Devriendt K., Tatton-Brown K., Pierpont M.E., Painter M., Worgan L., Reggin J., Hennekam R., Tsuchiya K., Pritchard C.C., Aracena M., Gripp K.W., Cordisco M., Van Esch H., Garavelli L., Curry C., Goriely A., Kayserilli H., Shendure J., Graham J J.r., Guerrini R, & Dobyns W.B. (2016). PIK3CA-associated developmental disorders exhibit distinct classes of mutations with variable expression and tissue distribution. JCI Insight, 1(9), e87623.
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4

Screening for PIK3R2 p.Gly373Arg Mutation

Check if the same lab product or an alternative is used in the 5 most similar protocols
To screen for the recurrent PIK3R2 mutation,
p.Gly373Arg, we performed locus-specific amplification of genomic DNA
followed by GS Junior sequencing. We designed fusion primers containing
genome-specific sequences along with distinct MIDs (multiplex identifier
sequences) used to differentiate samples being run together on the same
plate and sequencing adapters to generate amplicons ranging in size from 290
to 310 bp using primer3plus software. Primer sequences are available upon
request (Dr. Renzo Guerrini). Small DNA fragments were removed using
Agencourt AMPure XP (Beckman Coulter, Beverly, MA) according to the
manufacturer’s protocol. All amplicons were quantified using the
Quant-iT PicoGreen dsDNA reagent (Invitrogen Corporation, Life Technologies,
Carlsbad, CA), pooled at equimolar ratios, amplified by emulsion PCR using
the GS Junior Titanium emPCR kit (Lib-A kit, Roche Applied Science,
Mannheim, Germany) and pyrosequenced in the sense and antisense strands on a
GS Junior sequencer (Roche) following the manufacturer’s
instructions. We performed data analysis using the GS Amplicon Variant
Analyzer version 3.0 (AVAv3.0) software (Roche).
Mirzaa G., Conti V., Timms A.E., Smyser C.D., Ahmed S., Carter M., Barnett S., Hufnagel R.B., Goldstein A., Narumi-Kishimoto Y., Olds C., Collins S., Johnston K., Deleuze J.F., Nitschké P., Friend K., Harris C., Goetsch A., Martin B., Boyle E.A., Parrini E., Mei D., Tattini L., Slavotinek A., Blair E., Barnett C., Shendure J., Chelly J., Dobyns W.B, & Guerrini R. (2015). Characterization of mutations of the phosphoinositide-3-kinase regulatory subunit, PIK3R2, in perisylvian polymicrogyria: a next generation sequencing study. The Lancet. Neurology, 14(12), 1182-1195.
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