Original integration site analysis for K00339 performed in 2008 used a Southern blot and LAM-PCR approach. Briefly, DNA was extracted using the Puregene DNA Purification Kit (Quiagen, Inc., Valencia, CA, USA), followed by overnight digestion with BglII, EcoRI, or SacI. Samples were run on a 0.8% gel and transferred to a membrane. Next, samples were hybridized in Quick-Hyb solution (Amersham Biosciences Corp., Piscataway, NJ, USA) with 32P-labeled ires-GFP. Signals were detected using the Typhoon Phosphor Imaging System (Amersham Biosciences Corp., Piscataway, NJ, USA) LAM-PCR has been described previously [15 (link)] and involved the cloning of PCR products into the TOPO vector for sequencing. The collected sequences were aligned with the rhesus macaque genome assembly dated January 2006 from the University of Santa Cruz Genome Browser.
Typhoon phosphor imaging system
Manufactured by Cytiva
Sourced in United States
The Typhoon Phosphor Imaging System is a sensitive and versatile instrument designed for the detection and quantification of radioactive signals in gel-based and blotting applications. It utilizes a high-resolution laser scanning technology to capture digital images of phosphor-based samples.
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Lab products found in correlation
3 protocols using typhoon phosphor imaging system
1
Integration Site Analysis for K00339
2
mVPS35 Expression Analysis
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Total RNAs were extracted from hemibrains and subjected to Northern blot analysis using a 32P-labeled DNA probe (nucleotides 163–984 of mVPS35). Signal was detected using a Typhoon phosphor-imaging system (Amersham).
3
Northern Blotting of Yeast Genes
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Northern blotting was performed as described previously (11 (link)). Gene-specific probes were amplified by PCR from genomic DNA using oligonucleotide primers specific for ACT1, CAT1, and TRR1 (11 (link)). RNA levels were quantified using the Typhoon phosphorimaging system (Amersham Biosciences) and Image Quant software (GE Healthcare Life Sciences).
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