High Oligonucleotide Recovery From Liver Tissue
High Oligonucleotide Recovery From Liver Tissue
This is an Application Brief and does not contain a detailed Experimental section.
Abstract
This application brief provides guidance on the extraction of therapeutic oligonucleotides from tissue samples for quantitative analysis and demonstrates high oligonucleotide recovery from Liver Tissue.
Experimental
The following (Figure 1) summarizes the solvent-assisted tissue homogenization and oligonucleotide extraction workflow and protocol we use internally with our OligoWorks SPE Microplate Kit. Note we utilize a 1:10 ratio between the mg of tissue being homogenized and the total volumne (mL) of homogenization mix, including Rapizyme Proteinase K digestion reagents, buffer and solvent.
Figure 1. Oligonucleotide tissue extraction workflow and protocol using solvent assisted tissue homogenization and digestion with RapiZyme Proteinase K Digestion Module and OligoWorks SPE Microplate-2 mg/well.Note: equipment referenced is what we use in our lab, but alternate equipment with equivalent capabilities may be used instead.
LC-MS Analysis
Results and Discussion
Figure 2. Demonstration of OligoWorks SPE Microplate performance, with >70% oligonucleotide recovery using 0.05 g tissue/0.5 mL Proteinase K Digestion Module reagents (homogenized and digested 2 hrs at 55 °C, 600 rpm), and 100 µL of tissue supernatant purified using the OligoWorks SPE Microplate Kit, containing OligoWorks RapiZyme Proteinase K Digest Module and OligoWorks SPE Microplate-2 mg/well.
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720008270, April 2024
