LAMP2 and GAPDH Expression Analysis

To detect LAMP2 or GAPDH transcript expression, cellular RNA was extracted using RNeasy Mini Kit (Qiagen, Valencia, CA, United States) and cDNA was generated using the High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems, Foster City, CA, United States). Primers for LAMP2 and GAPDH amplification were described (Perez et al., 2016 (link)). LAMP2 cDNA was amplified using 2X ReddyMix PCR Master Mix (Thermo Fisher Scientific, Waltham, MA, United States) for 35 cycles. GAPDH cDNA was amplified for 30 cycles. PCR products were resolved by agarose gel.

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Pérez L., Sinn A.L., Sandusky G.E., Pollok K.E, & Blum J.S. (2018). Melanoma LAMP-2C Modulates Tumor Growth and Autophagy. Frontiers in Cell and Developmental Biology, 6, 101.

Publication 2018

RNeasy Mini Kit High-Capacity cDNA Reverse Transcription Kit 2X ReddyMix PCR Master Mix

Agarose Cdna Cellular Gapdh Lamp2 Primers Reverse transcription

Corresponding Organization : Indiana University – Purdue University Indianapolis

Other organizations : National Institute of Allergy and Infectious Diseases, National Institutes of Health, Indiana University Health

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Variable analysis

independent variables

Primers for LAMP2 and GAPDH amplification

dependent variables

LAMP2 transcript expression
GAPDH transcript expression

control variables

Cellular RNA extraction using RNeasy Mini Kit
CDNA generation using High-Capacity cDNA Reverse Transcription Kit
LAMP2 cDNA amplification for 35 cycles using 2X ReddyMix PCR Master Mix
GAPDH cDNA amplification for 30 cycles

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

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