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Questions related from Anita Ma
Trouble 1: I ran a PCR reaction and found a low molecular weight band (100 bp) when doing electrophoresis. 1ng plasmid (OD260/280=1.84) was used as template. Final concentration of primers was...
14 June 2024 5,489 6 View
I want to detect my target protein through WB and it worked well before. But now I could not get any signal for both my target and the GAPDH, so I ran SDS-PAGE. I expected many bands on the gel of...
24 May 2024 560 6 View
I want to copy a target gene from cDNA into a plasmid. The primers were designed according to the CDS sequence from NCBI. But when I performed PCR reactions I could not get any target bands. So...
16 April 2024 2,077 4 View
I found to use a wrong secondary Ab for my WB when I visualized it with ECL, can I wash with TBST for several times and incubate the right secondary Ab? Another question is why my bands are not...
17 February 2024 3,150 4 View
When I overexpressed a protein in a cell line, I found decrease of phospho-PLC (phospholipase C) at the protein level through WB. But when I knocked out the protein, there was no difference of...
11 January 2024 8,161 3 View
I want to use the Cre LoxP to knock out my target gene in a mammalian cell line. According to the mechanism, I need to add two loxP sequences at both ends of the gene through homologous...
22 November 2023 5,100 1 View
After proteomics analysis of my samples, a specific phosphorylation site on a kinase was found. But when I want to validate the result I could not find any commercial ab to that phosphorylated...
15 November 2023 348 6 View
This is my PCR 1.2% agarose gel result. I used a 25uL reaction system (0.3uM primers, 12.5uL Taq master mix, 1uL genome template and 5.5uL H2O). The primer Tm is 62℃, and reaction procedure is 94℃...
15 August 2023 1,370 5 View
I want to find out potential mechanisms of the effects of a certain gene on recombinant protein productivity . So proteomics analysis was performed between a high expression of my testing gene and...
28 March 2022 2,907 2 View
I am doing over expression of an endogenous target gene in a CHO cell line. When I did PCR (whole length of the gene) to verify the existence of my target gene, I could only see the gel band when...
28 October 2021 6,446 6 View
After transfection with the plasmid ( linearized ) and subcloning of the cell lines, RNA was extracted from the cell and then reverse transcripted to cDNA. When PCR reactions were run to verify...
26 February 2021 5,992 3 View
