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Questions related from Yen Teng Tai
I am using this BirAGFP mouse from the JAX lab (https://www.jax.org/strain/030420) and crossed it with my Nes-cre lines. I realized the GFP signal in the brain reduced as the mouse aged, has...
13 June 2023 6,805 1 View
Our mouse line carries the target protein tagged with BAP sequence and BirA protein that biotinylates BAP. We hope to improve the biotinylation efficiency by adding biotin to the diet, however, we...
18 October 2022 7,886 1 View
I am using cBirA mice that are designed to express BirA (biotin ligase) and EGFP following exposure to Cre recombinase. I crossed these mice with my own mice carrying BAP tag. I am wondering if I...
05 July 2022 3,077 0 View
We plan to perform a complementation assay in our human cerebral brain organoids by infecting our KO human embryonic stem cells with lentivirus that carries our gene of interest. Then, we will use...
07 February 2022 3,326 0 View
I extracted my brain organoid samples recently and ran them on SDS-PAGE. These brain organoid samples were coated with Matrigel, I tried to remove as many as I can by washing them in cold 1X PBS...
21 January 2022 8,549 4 View
I found that knockout of my gene of interest causes higher Wnt activity (as shown by TOP/FOP flash luciferase assay as well as RNA-seq) and hence, I am trying to figure out the mechanism that...
27 November 2021 4,470 3 View
Hi, I am currently trying to co-immunoprecipitate a protein but the interaction between that protein and its binding partner is not stable. Sometimes we able to detect its binding partner but...
30 November 2020 9,389 3 View
Has anyone tried to study Wnt signaling in HEK293T cells? I have been trying to measure the Wnt activity using TOP/FOP-FLASH assay but the readings that I got are rather similar in both TOP and...
29 August 2020 6,193 4 View
I have performed the ChIP and quantify the samples using Qubit Flurometer 2.0. Here are the results: Untagged control :2.72ng/mL Tagged sample: 3.17ng/mL I think the untagged strain should have...
10 April 2018 1,668 2 View
My protein A is being phosphorylated and I wish to know whether it is being phosphorylated by kinase B (a well-studied Serine/Threonine kinase). Is there any other methods other than doing...
06 April 2018 7,311 5 View
I need to prepare 1M of caffeine solution to be added into my liquid culture. We have anhydrous caffeine and caffeine sodium benzoate (40% of caffeine). For the anhydrous one, it is having...
10 August 2017 6,735 4 View
When I overexpressed a gene, it caused the cell growth to be severely affected and a large cell phenotype was observed. I am wondering is this normally happens when a protein is in excess? Or this...
08 July 2017 1,586 23 View
If overexpression of protein A can rescue the growth defect caused by mutation in protein B, can I consider protein A is working downstream of protein B? If yes, why is it so?
24 May 2017 6,149 23 View
I have a strain which my protein tagged with mNG and I wish to observe its expression using western blot. I wonder can I use GFP antibody to detect mNeonGreen signals? Do they have the same...
24 February 2017 2,417 7 View
I am preparing samples for BigDye sequencing. After PCR, I did ethanol precipitation to clean up the samples. In order to dry up the samples (after removing 70% ethanol), is it okay for me to dry...
25 October 2016 9,614 7 View
One of the proteins in S.pombe that I am working on is tagged with FLAG, so can I still call this strain as wild type as it contains a fragment of foreign DNA? If no, what is the right term for it?
02 June 2016 5,073 3 View
I have digested my PCR product with NEB Not1 and Nde1 for 1hr. I have cut the band and purified the product. However I afraid the digestion is incomplete, can I re-digest them again? Is there any...
24 May 2016 7,152 15 View
May I know 400U of lambda phosphatase can dephosphorylate how many microgram of protein? I am using lambda phosphatase from Cosmo Bio.
29 February 2016 5,130 4 View
May I know what is the maximum number of plasmids (with different origins of replication) can be taken up by a cell? What factors affect this? The maximum I have tried was two.
23 February 2016 8,689 4 View
I have ran 16S PCR on my metagenomic DNA samples (extracted from wastewater). However, I got multiple bands. Is this normal?
17 June 2015 1,480 18 View
The size of my gene is 1.7kb plus promoter is 800bp, so when I do PCR using plasmid primers, I should get 2300bp. However, I only can get 1500bp after I ran gel. I have screened through many...
22 July 2014 7,865 19 View
I have sent my soil sample for amplicon sequencing, the primers are targeted on bacteria V3-V5 region. However, the result that I obtained contains other things such as animalia, archae and...
03 December 2013 469 5 View
After I did a plasmid miniprep using Qiagen kit, the band is smeared and some even show very a low yield. May I know why?
27 October 2013 2,673 1 View
After I did blue-white screening, I picked the white colonies and subculture on LB/Amp plate. They can grow well but when I subculture again on new plate, they cannot grow. May I know the reason?...
24 October 2013 7,258 2 View
May I know if it is true that gene with any sequence also can be inserted into pGEM-T Easy? Since it has AT-overhang, so will this affect the type of gene inserted?
09 October 2013 1,495 15 View
I am extracting DNA from soil sample based on Zhou's conventional method but I am facing problems with the humic acids. The humic acids will decrease the purity of DNA and thus, inhibits the...
29 April 2013 8,970 3 View
the sample using gel electrophoresis. After that, I sonicate the sample but when I want to cut gel
25 April 2013 4,935 10 View