For evaluation of liver parasite burden, hepatocytes were extracted with a 30-minute liver digestion, mashing through 100 αM cell strainers, and a hepatocyte spin (55 g, 3 minutes, low brake). Pellets were resuspended in TRIzol (Ambion) and flash frozen in liquid nitrogen. RNA was extracted and purified from the pellets using the RNA Clean & Concentrator kit (Zymo Research, #R1018) according to the manufacturer instruction. RNA samples were amplified and quantified using Taq polymerase (Applied Biosystems, #4444427) Pb 18S rRNA forward and reverse primers and probe (Integrated DNA Technologies, #142265234, 142265235, 115842913), mouse GAPDH forward and reverse primers and probe (Applied Biosystems, #4308313) the University of Iowa Genomics Core facility (7900 HT). Pb 18S rRNA transcript counts were normalized to mouse GAPDH counts.
Pb 18s rrna forward and reverse primers and probe
Manufactured by Integrated DNA Technologies
The Pb 18S rRNA forward and reverse primers and probe are designed to target and amplify a specific region of the 18S ribosomal RNA (rRNA) gene of the Plasmodium berghei parasite. These tools are intended for use in molecular biology research applications that require the detection and quantification of P. berghei 18S rRNA sequences.
Automatically generated - may contain errors
2 protocols using pb 18s rrna forward and reverse primers and probe
1
Quantifying Liver Parasite Burden
2
Quantifying Liver Parasite Burden
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For evaluation of liver parasite burden, hepatocytes were extracted with a 30-minute liver digestion, mashing through 100 αM cell strainers, and a hepatocyte spin (55 g, 3 minutes, low brake). Pellets were resuspended in TRIzol (Ambion) and flash frozen in liquid nitrogen. RNA was extracted and purified from the pellets using the RNA Clean & Concentrator kit (Zymo Research, #R1018) according to the manufacturer instruction. RNA samples were amplified and quantified using Taq polymerase (Applied Biosystems, #4444427) Pb 18S rRNA forward and reverse primers and probe (Integrated DNA Technologies, #142265234, 142265235, 115842913), mouse GAPDH forward and reverse primers and probe (Applied Biosystems, #4308313) the University of Iowa Genomics Core facility (7900 HT). Pb 18S rRNA transcript counts were normalized to mouse GAPDH counts.
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