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Cf384 touch real time prc detection system

Manufactured by Bio-Rad
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The CF384 Touch Real-Time PCR Detection System is a high-throughput real-time PCR instrument. It features a 384-well thermal cycler and can perform quantitative and qualitative real-time PCR analysis.

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Lab products found in correlation

Cdna archive kit, by Thermo Fisher Scientific (4 mentions) Rneasy mini kit, by Qiagen (4 mentions) Anti cd11c macs beads, by Miltenyi Biotec (2 mentions) Deoxyribonuclease 1, by Worthington (2 mentions) Qiashredder column, by Qiagen (2 mentions) Collagenase type 4, by Worthington (2 mentions) Rneasy cleanup kit, by Qiagen (2 mentions) Trizol reagent, by Thermo Fisher Scientific (2 mentions) Rnalater, by Qiagen (2 mentions)

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Real-Time System

4 protocols using cf384 touch real time prc detection system

1

Splenic DC Isolation and Gene Expression

Cited 1 time
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As previously described (Stanley et al., 2008 (link)) mice were infected with L. donovani and sacrificed 5 hours later. Spleens were digested in collagenase type IV (1 mg/ml; Worthington, Lakewood, NJ) and deoxyribonuclease I (0.5 mg/ml; Worthington) at RT for 45 minutes. Splenocytes were processed as described above. Splenic DCs were isolated from splenocyte preparations by positive selection with anti-CD11c MACS beads, according to the manufacturer’s instructions (Miltenyi Biotec). DCs were also stained for MHCII and CD86 and analyzed by flow cytometry, as described above. Purified DCs were stored in buffer RLT and homogenized in QIAshredder columns (both from QIAGEN). Total RNA was extracted from purified DC using the RNeasy Mini Kit with on-column DNase digestion (both from QIAGEN). RNA samples were reverse transcribed into cDNA using the cDNA Archive Kit (Applied Biosystems, Foster City, CA). RT-qPCR for Il12 and Il10 was performed on CF384 Touch Real-Time PRC Detection System (BIO-RAD) using the TaqMan Gene Expression Assay (Applied Bioscience). Relative quantification was performed using the comparative CT method relative to Hprt and beta2m.
Kumar R., Bunn P.T., Singh S.S., Ng S.S., de Oca M.M., De Labastida Rivera F., Chauhan S.B., Singh N., Faleiro R.J., Edwards C.L., Frame T.C., Sheel M., Austin R.J., Lane S.W., Bald T., Smyth M.J., Hill G.R., Best S.E., Haque A., Corvino D., Waddell N., Koufariotis L., Mukhopadhay P., Rai M., Chakravarty J., Singh O.P., Sacks D., Nylen S., Uzonna J., Sundar S, & Engwerda C.R. (2020). Type I Interferons Suppress Anti-parasitic Immunity and Can Be Targeted to Improve Treatment of Visceral Leishmaniasis. Cell reports, 30(8), 2512-2525.e9.
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2

Splenic DC Isolation and Gene Expression

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
As previously described (Stanley et al., 2008 (link)) mice were infected with L. donovani and sacrificed 5 hours later. Spleens were digested in collagenase type IV (1 mg/ml; Worthington, Lakewood, NJ) and deoxyribonuclease I (0.5 mg/ml; Worthington) at RT for 45 minutes. Splenocytes were processed as described above. Splenic DCs were isolated from splenocyte preparations by positive selection with anti-CD11c MACS beads, according to the manufacturer’s instructions (Miltenyi Biotec). DCs were also stained for MHCII and CD86 and analyzed by flow cytometry, as described above. Purified DCs were stored in buffer RLT and homogenized in QIAshredder columns (both from QIAGEN). Total RNA was extracted from purified DC using the RNeasy Mini Kit with on-column DNase digestion (both from QIAGEN). RNA samples were reverse transcribed into cDNA using the cDNA Archive Kit (Applied Biosystems, Foster City, CA). RT-qPCR for Il12 and Il10 was performed on CF384 Touch Real-Time PRC Detection System (BIO-RAD) using the TaqMan Gene Expression Assay (Applied Bioscience). Relative quantification was performed using the comparative CT method relative to Hprt and beta2m.
Kumar R., Bunn P.T., Singh S.S., Ng S.S., de Oca M.M., De Labastida Rivera F., Chauhan S.B., Singh N., Faleiro R.J., Edwards C.L., Frame T.C., Sheel M., Austin R.J., Lane S.W., Bald T., Smyth M.J., Hill G.R., Best S.E., Haque A., Corvino D., Waddell N., Koufariotis L., Mukhopadhay P., Rai M., Chakravarty J., Singh O.P., Sacks D., Nylen S., Uzonna J., Sundar S, & Engwerda C.R. (2020). Type I Interferons Suppress Anti-parasitic Immunity and Can Be Targeted to Improve Treatment of Visceral Leishmaniasis. Cell reports, 30(8), 2512-2525.e9.
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3

Murine Spleen and Liver RNA Extraction

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Pieces of murine spleen and liver were collected into RNAlater (QIAGEN) and total RNA was extracted using TRIzol Reagent (Invitrogen Life Technologies), RNeasy Cleanup Kit and a RNeasy Mini Kit with on-column DNase digestion (all from QIAGEN) as per the manufacturer’s instructions. RNA samples were reverse transcribed into cDNA using the cDNA Archive Kit (Applied Biosystems, Foster City, CA). RT-qPCR for Ifng, Tnf and Il10 was performed on CF384 Touch Real-Time PRC Detection System (BIO-RAD) using the TaqMan Gene Expression Assay (Applied Biosciences). Relative quantification was performed using the comparative CT method relative to Hprt and beta2m.
Kumar R., Bunn P.T., Singh S.S., Ng S.S., de Oca M.M., De Labastida Rivera F., Chauhan S.B., Singh N., Faleiro R.J., Edwards C.L., Frame T.C., Sheel M., Austin R.J., Lane S.W., Bald T., Smyth M.J., Hill G.R., Best S.E., Haque A., Corvino D., Waddell N., Koufariotis L., Mukhopadhay P., Rai M., Chakravarty J., Singh O.P., Sacks D., Nylen S., Uzonna J., Sundar S, & Engwerda C.R. (2020). Type I Interferons Suppress Anti-parasitic Immunity and Can Be Targeted to Improve Treatment of Visceral Leishmaniasis. Cell reports, 30(8), 2512-2525.e9.
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4

Murine Spleen and Liver RNA Extraction

Check if the same lab product or an alternative is used in the 5 most similar protocols
Pieces of murine spleen and liver were collected into RNAlater (QIAGEN) and total RNA was extracted using TRIzol Reagent (Invitrogen Life Technologies), RNeasy Cleanup Kit and a RNeasy Mini Kit with on-column DNase digestion (all from QIAGEN) as per the manufacturer’s instructions. RNA samples were reverse transcribed into cDNA using the cDNA Archive Kit (Applied Biosystems, Foster City, CA). RT-qPCR for Ifng, Tnf and Il10 was performed on CF384 Touch Real-Time PRC Detection System (BIO-RAD) using the TaqMan Gene Expression Assay (Applied Biosciences). Relative quantification was performed using the comparative CT method relative to Hprt and beta2m.
Kumar R., Bunn P.T., Singh S.S., Ng S.S., de Oca M.M., De Labastida Rivera F., Chauhan S.B., Singh N., Faleiro R.J., Edwards C.L., Frame T.C., Sheel M., Austin R.J., Lane S.W., Bald T., Smyth M.J., Hill G.R., Best S.E., Haque A., Corvino D., Waddell N., Koufariotis L., Mukhopadhay P., Rai M., Chakravarty J., Singh O.P., Sacks D., Nylen S., Uzonna J., Sundar S, & Engwerda C.R. (2020). Type I Interferons Suppress Anti-parasitic Immunity and Can Be Targeted to Improve Treatment of Visceral Leishmaniasis. Cell reports, 30(8), 2512-2525.e9.
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