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Mini janus

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The Mini Janus is a compact, benchtop liquid handling workstation designed for automated pipetting and sample preparation. It provides precise and accurate liquid handling for a variety of applications in life science research and clinical laboratories.

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Lab products found in correlation

Truseq rna sample preparation protocol, by Illumina (3 mentions) Quant it ribogreen rna assay kit, by Thermo Fisher Scientific (3 mentions) Kapa library quantification kit for sequencing platforms, by Illumina (2 mentions) Hybridization buffer, by Illumina (2 mentions) Minijanus, by Agilent Technologies (2 mentions) Fibroblast growth factor 2 (fgf2), by Merck Group (1 mentions) Celltiter glo luminescent cell viability assay, by Promega (1 mentions) Bravo liquid handling platform, by Agilent Technologies (1 mentions) Bravo platform, by Agilent Technologies (1 mentions) Kinase inhibitor library, by Enzo Life Sciences (1 mentions)

Spelling variants of this product

Janus MDT Mini (2 citations)

7 protocols using mini janus

1

RNA-Seq Analysis of FGF2, Fulvestrant, and Palbociclib Treatments

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CAMA1 cells were plated in full media ± 2 ng/mL FGF2 (Sigma) and then treated with DMSO or 1 μM fulvestrant plus 1 μM palbociclib ± 1 μM lucitanib for 6 h. Cells were harvested and RNA was purified using a RNA purification kit (Maxwell, Promega). Total RNA was quantified using the Quant-iT Ribo-Green RNA Assay Kit (Invitrogen) and normalized to 4 ng/mL; 200 ng of each sample were used for library preparation in an automated variant of the Illumina Tru Seq RNA Sample Preparation protocol (Revision A, 2010). This method uses oligo(dT) beads to select mRNA from the total RNA sample and is followed by heat fragmentation and cDNA synthesis from the RNA template. The resultant cDNA went through library preparation (end repair, base “A” addition, adapter ligation, and enrichment) using Broad Institute–designed indexed adapters for multiplexing. After enrichment, libraries were quantitated with qPCR using the KAPA LibraryQuantification Kit for Illumina Sequencing Platforms and pooled equimolarly. The entire process was performed in a 96-well format with all pipetting done by either the Agilent Bravo or PerkinElmer JANUS Mini liquid handlers.
Formisano L., Lu Y., Servetto A., Hanker A.B., Jansen V.M., Bauer J.A., Sudhan D.R., Guerrero-Zotano A.L., Croessmann S., Guo Y., Ericsson P.G., Lee K.M., Nixon M.J., Schwarz L.J., Sanders M.E., Dugger T.C., Cruz M.R., Behdad A., Cristofanilli M., Bardia A., O’Shaughnessy J., Nagy R.J., Lanman R.B., Solovieff N., He W., Miller M., Su F., Shyr Y., Mayer I.A., Balko J.M, & Arteaga C.L. (2019). Aberrant FGFR signaling mediates resistance to CDK4/6 inhibitors in ER+ breast cancer. Nature Communications, 10, 1373.
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2

Automated mRNA-Seq Library Preparation

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Total RNA was quantified using the Quant-iT™ RiboGreen® RNA Assay Kit (Invitrogen) and normalized to 4 ng/μL; 200 ng of each sample were used for library preparation on an automated variant of the Illumina TruSeq™ RNA Sample Preparation protocol (Revision A, 2010). This method uses oligo(dT) beads to select mRNA from the total RNA sample and is followed by heat fragmentation and cDNA synthesis from the RNA template. The resultant cDNA then goes through library preparation (end repair, base ‘A’ addition, adapter ligation and enrichment) using Broad Institute-designed indexed adapters for multiplexing. After enrichment, the libraries were quantified with qPCR using the KAPA Library Quantification Kit for Illumina Sequencing Platforms and then pooled equimolarly. The entire process is performed in a 96-well format and all pipetting is done by either Agilent Bravo or PerkinElmer JANUS Mini liquid handlers.
Giltnane J.M., Hutchinson K.E., Stricker T.P., Formisano L., Young C.D., Estrada M.V., Nixon M.J., Du L., Sanchez V., Ericsson P.G., Kuba M.G., Sanders M.E., Mu X.J., Van Allen E.M., Wagle N., Mayer I., Abramson V., Gómez H., Rizzo M., Toy W., Chandarlapaty S., Mayer E.L., Christiansen J., Murphy D., Fitzgerald K., Wang K., Ross J.S., Miller V.A., Stephens P.J., Yelensky R., Garraway L., Shyr Y., Meszoely I., Balko J.M, & Arteaga C.L. (2017). Genomic profiling of ER+ breast cancers after short-term estrogen suppression reveals alterations associated with endocrine resistance. Science translational medicine, 9(402), eaai7993.
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3

Protocol for RNA extraction and sequencing

Cited 1 time
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Core biopsies were flash frozen in liquid N2 and stored at −80°C until RNA extraction was performed as described elsewhere (26 (link)). Total RNA was quantified using the Quant-iT™ RiboGreen® RNA Assay Kit (Invitrogen) and normalized to 4 ng/μL; 200 ng of each sample were used for library preparation in an automated variant of the Illumina Tru Seq™ RNA Sample Preparation protocol (Revision A, 2010). This method uses oligo(dT) beads to select mRNA from the total RNA sample and is followed by heat fragmentation and cDNA synthesis from the RNA template. The resultant cDNA went through library preparation (end repair, base ‘A’ addition, adapter ligation, and enrichment) using Broad Institute-designed indexed adapters for multiplexing. After enrichment, libraries were quantitated with qPCR using the KAPA Library Quantification Kit for Illumina Sequencing Platforms and pooled equimolarly. The entire process was performed in a 96-well format with all pipetting done by either the Agilent Bravo or PerkinElmer JANUS Mini liquid handlers.
Formisano L., Stauffer K.M., Young C.D., Bhola N.E., Guerrero A.L., Jansen V.M., Estrada M.V., Hutchinson K.E., Giltnane J.M., Schwarz L.J., Lu Y., Balko J.M., Deas O., Cairo S., Judde J.G., Mayer I.A., Sanders M.E., Dugger T.C., Bianco R., Stricker T.P, & Arteaga C.L. (2017). Association of FGFR1 with ERα maintains ligand-independent ER transcription and mediates resistance to estrogen deprivation in ER+ breast cancer. Clinical cancer research : an official journal of the American Association for Cancer Research, 23(20), 6138-6150.
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4

Screening Combination Therapies for HCC

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HCC PDXOs and THLE-2 cells were seeded at 2000 cells/well density in 384-well plates 1 day before drug treatment. The drug combinations used for QPOP for nine drugs at three dosage levels were generated using an orthogonal array composite design (OACD) as described by Xu et al. [42 ], which combines a three-level orthogonal array with a two-level fractional factorial design to achieve the least number of combinations for factor screening. The three dosage levels used are IC0, IC10, and IC20, which were determined based on the dose-response curves of each drug across the panel of HCC PDXOs. Drug combinations were prepared using an automated liquid handler (Mini Janus, Perkin Elmer), and after 48 hours of drug treatment, cell viability was measured using CellTiter-Glo Luminescent Cell Viability Assay (Promega, G5771) as per manufacturer’s instructions. QPOP analysis was performed as previously described [31 (link)], using the viabilities of THLE-2 (viabilityTHLE2) and HCC PDXOs (viabilityPDXO) as readouts. Briefly, the viability for each experimental combination was used as data points to fit a second-order quadratic series that generates all possible drug combinations with their corresponding projected output values. All two-drug combinations were subsequently ranked according to the output and used for hierarchical clustering and frequency analysis.
Lim J.J., Hooi L., Dan Y.Y., Bonney G.K., Zhou L., Chow P.K., Chee C.E., Toh T.B, & Chow E.K. (2022). Rational drug combination design in patient-derived avatars reveals effective inhibition of hepatocellular carcinoma with proteasome and CDK inhibitors. Journal of Experimental & Clinical Cancer Research : CR, 41, 249.
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5

Library Quantification and Pooling

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After post-capture enrichment, libraries were quantified using PicoGreen, normalized to equal concentration using a Perkin Elmer MiniJanus instrument, and pooled by equal volume on the Agilent Bravo platform. Library pools were then quantified using quantitative PCR (KAPA Biosystems) with probes specific to the ends of the adapters; this assay was automated using Agilent’s Bravo liquid handling platform. On the basis of quantitative PCR (qPCR) quantification, libraries were brought to 2 nM and denatured using 0.2 N NaOH on the Perkin-Elmer MiniJanus. After denaturation, libraries were diluted to 20 pM using hybridization buffer purchased from Illumina.
Taylor-Weiner A., Zack T., O’Donnell E., Guerriero J.L., Bernard B., Reddy A., Han G.C., AlDubayan S., Amin-Mansour A., Schumacher S.E., Litchfield K., Turnbull C., Gabriel S., Beroukhim R., Getz G., Carter S.L., Hirsch M.S., Letai A., Sweeney C, & Van Allen E.M. (2016). Genomic evolution and chemoresistance in germ-cell tumours. Nature, 540(7631), 114-118.
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6

Automated Library Quantification and Normalization

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After post-capture enrichment, libraries were quantified using PicoGreen, normalized to an equal concentration using a Perkin Elmer MiniJanus instrument, and pooled by equal volume on the Agilent Bravo platform. Library pools were then quantified using quantitative PCR (KAPA Biosystems) with probes specific to the ends of the adapters; this assay was automated using Agilent's Bravo liquid handling platform. Based on qPCR quantification, libraries were brought to 2 nM and denatured using 0.2 N NaOH on the Perkin-Elmer MiniJanus. After denaturation, libraries were diluted to 20 pM using hybridization buffer purchased from Illumina.
Giltnane J.M., Hutchinson K.E., Stricker T.P., Formisano L., Young C.D., Estrada M.V., Nixon M.J., Du L., Sanchez V., Ericsson P.G., Kuba M.G., Sanders M.E., Mu X.J., Van Allen E.M., Wagle N., Mayer I., Abramson V., Gómez H., Rizzo M., Toy W., Chandarlapaty S., Mayer E.L., Christiansen J., Murphy D., Fitzgerald K., Wang K., Ross J.S., Miller V.A., Stephens P.J., Yelensky R., Garraway L., Shyr Y., Meszoely I., Balko J.M, & Arteaga C.L. (2017). Genomic profiling of ER+ breast cancers after short-term estrogen suppression reveals alterations associated with endocrine resistance. Science translational medicine, 9(402), eaai7993.
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7

Kinase Inhibitor Drug Screening

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Hep3B and THLE-2 cells were dissociated with trypsin and seeded into 96-well plates at a density of 5000 cells per well. Cells were allowed to adhere and recover for 24 h prior to drug treatment. Thereafter, cells were subjected to 1 µM and 5 µM of each drug from the kinase inhibitor library (Enzo Life Sciences, Farmingdale, NY). The drug treatment was executed by the liquid-handling system Mini Janus (PerkinElmer, Waltham, MA). The drugs were diluted in media to a concentration of 20 µM before dispensing the drugs into master plates containing the respective media to a final concentration of 1 µM by the Mini Janus system (Fig. 1B). Thereafter, the library of drugs at 1 µM was added to the plates containing the respective cells and incubated for 24 h.
Mohd Abdul Rashid M.B., Toh T.B., Silva A., Nurrul Abdullah L., Ho C.M., Ho D, & Chow E.K. (2015). Identification and Optimization of Combinatorial Glucose Metabolism Inhibitors in Hepatocellular Carcinomas. Journal of laboratory automation, 20(4).
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