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Maximah rt

Manufactured by New England Biolabs
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MaximaH RT is a reverse transcriptase enzyme used for the synthesis of first-strand cDNA from RNA templates. It is suitable for a variety of applications, including RT-PCR, cDNA library construction, and gene expression analysis.

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Lab products found in correlation

Monarch total rna extraction kit, by New England Biolabs (2 mentions) Ariamx qpcr machine, by Agilent Technologies (2 mentions) Aria 1, by Agilent Technologies (2 mentions) Fitcanti hla dq, by BioLegend (2 mentions) Taqman master mix, by Thermo Fisher Scientific (2 mentions)

2 protocols using maximah rt

1

HLA-DQB1 Expression Analysis in Clones

Check if the same lab product or an alternative is used in the 5 most similar protocols
For analysis of HLA-DQB1 expression on HH WT (C/C) and
ALT (G/G) clones, RNA was extracted from clones using a Monarch Total RNA
extraction kit (NEB). cDNA was synthesized using MaximaH RT (NEB) enzyme
following manufacturer’s protocol and oligoDT primers. cDNA was diluted 1
in 4 with HLA-DQB1 (Assay ID:Hs00409790) and
actinB (Assay ID:Hs01060665) probes and Taqman MasterMix
(Thermofisher). Samples were run on an ARIAmx qPCR machine (Agilent) and data
analyzed with Aria 1.5 (Agilent) software. Expression is represented as
2-delta(HLA-DQB1 Ct - ActinBCt). For analysis of protein expression, clones that survived after
3–4 months of culture were washed with PBS and stained with FITC
anti-HLA-DQ (Biolegend, Clone: HLADQ1) for 30 minutes on ice and cell surface
expression assessed by flow cytometry. Data was analyzed using Flowjo and
Graphpad PRISM.
Gutierrez-Arcelus M., Baglaenko Y., Arora J., Hannes S., Luo Y., Amariuta T., Teslovich N., Rao D.A., Ermann J., Jonsson A.H., Navarrete C., Rich S.S., Taylor K.D., Rotter J.I., Gregersen P.K., Esko T., Brenner M.B, & Raychaudhuri S. (2020). Allele-specific expression changes dynamically during T cell activation in HLA and other autoimmune loci. Nature genetics, 52(3), 247-253.
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2

HLA-DQB1 Expression Analysis in Clones

Check if the same lab product or an alternative is used in the 5 most similar protocols
For analysis of HLA-DQB1 expression on HH WT (C/C) and
ALT (G/G) clones, RNA was extracted from clones using a Monarch Total RNA
extraction kit (NEB). cDNA was synthesized using MaximaH RT (NEB) enzyme
following manufacturer’s protocol and oligoDT primers. cDNA was diluted 1
in 4 with HLA-DQB1 (Assay ID:Hs00409790) and
actinB (Assay ID:Hs01060665) probes and Taqman MasterMix
(Thermofisher). Samples were run on an ARIAmx qPCR machine (Agilent) and data
analyzed with Aria 1.5 (Agilent) software. Expression is represented as
2-delta(HLA-DQB1 Ct - ActinBCt). For analysis of protein expression, clones that survived after
3–4 months of culture were washed with PBS and stained with FITC
anti-HLA-DQ (Biolegend, Clone: HLADQ1) for 30 minutes on ice and cell surface
expression assessed by flow cytometry. Data was analyzed using Flowjo and
Graphpad PRISM.
Gutierrez-Arcelus M., Baglaenko Y., Arora J., Hannes S., Luo Y., Amariuta T., Teslovich N., Rao D.A., Ermann J., Jonsson A.H., Navarrete C., Rich S.S., Taylor K.D., Rotter J.I., Gregersen P.K., Esko T., Brenner M.B, & Raychaudhuri S. (2020). Allele-specific expression changes dynamically during T cell activation in HLA and other autoimmune loci. Nature genetics, 52(3), 247-253.
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