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Questions related from Igor Bychkov
Hello dear colleagues. I am transfecting 10^6 293LTV cells in 6-well plates with lentiviral plasmids for virus production. I am doing calcium phosphate transfection with 5 mkg of DNA and homemade...
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Hello dear colleagues. I have a relatively frequent variant (MAF ~ 3% in general population). Using minigene assay i found out that it leads to some amount of exon skipping and subsecuent nonsense...
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Hello dear colleagues, i did the site directed mutagenesis of the plasmid with Thermo Scientific Phusion Site-Directed Mutagenesis Kit to introduce a point mutation with phosphorylated primers....
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Hello dear colleagues! Im looking for a simple method to remove platelets from blood samples. Gradient centrifugation doesnt work well because platelets have same density as mononuclear cells, and...
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Hello dear colleagues. Im trying to completely eliminate platelets from blood samples and found a method with using serum tubes for blood collection followed by ficoll centrifugation. I tried it...
02 June 2018 5,201 3 View
Hello dear collegues! Im using gradient centrifugation for isoltation of PBMC from blood samples. 5ml of fresh blood + 5ml of ficoll 1077 at 400g for 30min, 18C. I remove plasma layer and collect...
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Hello dear collegues. Im using sybr green 1 qpcr assay for evaluation of relative mitochondrial copy numbers in dna samples from human blood. Im using primers for nuclear and mitochondrial genome...
11 November 2017 725 4 View
Hello dear colleagues. Im trying to clone my dna fragment (2000 bp) using TA vector and E.Coli cells. In PCR step from cdna template i was using high sensitivity polymerase with 5`-3` proofreading...
30 October 2017 3,669 4 View
Hello dear colleagues. Im doing qPCR research of mitochondrial copy number in DNA samples from blood. Im using StepOne plus thermocycler and Applied biosystems 0.1 mL 96-well plates. While doing...
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