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NeoGeneStar 100 Randolph Road, Suite 2B Somerset, NJ 08873 Tel: (732) 421-4567 Fax (908) 756-4483 www.NeoGeneStar.com NeoGeneStarTM Spike Recovery Protocol For determination of recovery of cfDNA from plasma, serum or urine. This protocol serves as a positive control for validation of the cfDNA purification. Suggested DNA ladders: 50 BP DNA Ladder Plus ITSI Biosciences GeneRuler™ 50 bp DNA Ladder Low Range DNA Ladder M-202 Thermo Fisher Scientific SM0371 Jena BioScience Diamond DNA Ladder 50-150bp Rockland MB-206-0500 Recommend spike 300 ng of ladder added to plasma or urine prior to addition of lysis/binding buffer. Proceed with cfDNA purification as usual. Recovery should be >90% which can be measured with Qubit, BioAnalyzer (electropherogram) and/or gel electrophoresis. For Gel Electrophoresis – load 1 lane with appropriate amount of marker, ie. If cfDNA is recovered in 30ul and 10ul is used for gel analysis, the control marker lane should have 100ng of DNA. Typical gel image showing >90% recovery: Lane 100 ng 100 bp DNA Ladder 2 Recovery of 100 bp DNA Ladder from 5ml of EDTA human Plasma Notes: 2 3 Contents 1 3 1 Recovery of 100 bp DNA Ladder from 5ml of human urine 4% E-Gel EX Page 1 of 1 - 500 bp 400 bp 300 bp - 200 bp - 100 bp Life Technologies NeoGeneStarTM Spike Recovery Positive Control Protocol 09-15-2015
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