29 Questions 59 Answers 0 Followers
Questions related from Shuaichen Liu
Hi researchers, As the title shows, recently I made a construct with a 8xHis tag at N ternimal of HBx (154 aa). The voctor is pCDNA3.1+ and the sequencing result showed everything was correct. I...
11 November 2017 6,463 5 View
Hi researchers, I'm using a PCR mycoplasma detection protocol which is found online and it works well for me. Link as...
10 October 2017 4,357 4 View
Hi all, I'm doing a de novo protein structure prediction using https://zhanglab.ccmb.med.umich.edu There are two modes for the prediction. QUARK for less than 200 AA and I-TARSSER for bigger...
08 August 2017 2,252 2 View
Hi there, I want to treat several liver cell lines with IL-6 and then check some genes' expression at mRNA level using real time PCR. Is there any recommendation for positive control in this...
07 July 2017 6,013 3 View
Hi researchers, I want to over express a protein in Hep3B cell line. The expressing vector I use is attached here. The packaging vectors are pMD2.G, pRSV-Rev and pMDLg/pRRE. The amount I use for...
07 July 2017 1,704 4 View
Hi dear researchers, I'm checking a table for a antibody array result. For each protein the company detect, they give a Swiss prot ID which can be checked at http://www.uniprot.org/. I want to ask...
04 April 2017 2,343 3 View
Hi researchers who study BMP2, I'm doing western blot for BMP2 recently and really need some help from those who ever done western for this protein! My major question is what is the real size for...
01 January 2017 7,388 4 View
Hi researchers, I made a construct with a Flag tagged to the N-terminal of my aim protein. I've already done the lentivirus infection and selected for over 2 weeks. I did a real time PCR to check...
01 January 2017 6,952 7 View
Hi all, I want to order one siRNA from one publication. In the publication, they used 3 siRNAs and I want to order the most efficient one first. This is a 19nt siRNA. But when I do the blast,...
10 October 2016 452 4 View
I want check the protein level of BMP2 (Bone morphogenetic protein 2). I see someone use cell lysiate to do western blot while others use cell culture supernantant to do ELISA for BMP2. I know...
09 September 2016 1,729 4 View
Hi researchers, When I do primer blast, the result will show me an ID of the gene that my primer will bind to. If I'm not wrong, NM_xxxxxxxxxx means it's a human sequence. Then what does...
09 September 2016 4,182 2 View
Hi researchers, I try to find a pair of primers for CXCL12 from publications. One strange thing I found was that, we I use the primers from publication to do blast in NCBI primer tool, they cannot...
09 September 2016 3,885 3 View
Hi researchers, I want to look at the mRNA expression data for a certain cell line (if someone has done it) in public database, how can I do that? For example, if I want to check HepG2.2.15 cell...
08 August 2016 1,326 5 View
Hi researchers, I got one tube of frozen BxPC-3 cell from my colleague and thawed it yesterday. My colleague told me when he cultured this cell previously, a lot of them will float in the flask...
08 August 2016 3,939 0 View
Hi researchers, I want to measure IL-6 secreted by cultured cells at different time point, using the supernantant. For example 12, 24, 48h after stimuli. But one thing I realize is that the ELISA...
07 July 2016 2,187 3 View
Hi researchers, I want to ask a question about ELISA dilution factor with my data. For the same sample, the first time I used 1:2 dilution (1 plasma + 1 diluent) and the OD value was 2.482. The...
07 July 2016 5,302 3 View
Hi researchers, I want to see whether chronic hepatitis B infection will increase the expression of one gene. I'm not sure how to “generate” this condition. Can I use just inflammation cytokines...
07 July 2016 789 4 View
Hi researchers, I'm doing PCR from a 8kb fragment from genomic DNA. Since I need to do many samples, I find that the template quality seems affect the yield. Can see the attached figure. Just...
05 May 2016 8,623 4 View
Hi researchers, Please see the attached image. This is a gel image of a 8kb PCR product from genomic.Each two adjacent lanes are the same sample from a 50ul PCR system. I use 6 pairs of primers to...
04 April 2016 1,541 7 View
Hi researchers, Is it necessary to have a negative control for ELISA? I see someone mentions that negative control is serum without my aim protein. If it is so, how can I get such kind of...
04 April 2016 8,839 8 View
Hi dear researchers, I'm going to use ELISA to check a protein level in serum. I bought the ELISA plate coated with the antibody and I can get the protein concentration just by this plate. What I...
04 April 2016 6,749 8 View
Hi researchers, My colleague passed me same serum samples (300ul each tube). He thawed it from stock and his operation was normative. I wanted to aliquot 50ul from each tube but I didn't know I...
04 April 2016 212 18 View
Hi researchers, Now I need to do end repair and dA-tailing for a 8000bp DNA fragment. My lab has NEBNext End Repair Module(E6050) and dA-tailing Module(E6053). But when I read the protocol, both...
04 April 2016 5,800 2 View
Hi dear researchers, Now I'm using the MinION to sequence a long PCR product. I realize that maybe during the end repair process, I lost a lot of my DNA sample, because there is an obvious...
03 March 2016 8,482 16 View
Hi everyone, I want to add some barcode sequences to primers then PCR them from different samples. Just want to know if there are any principles behind the barcode? I mean the length or the...
02 February 2016 8,705 2 View
Hi dear researchers, Just want to ask whether you are using Minion for sequencing. I'm going to sequence a whole gene (~8kb) with it. If I want to combine a few samples and sequence together in...
02 February 2016 3,398 4 View
I tried quite a few times to PCR a ~8kb fragment from genomic DNA but cannot get it. Here I just show the newest trials and really hope someone can give me some advice. 1.The first attached file...
02 February 2016 8,149 13 View
When I'm designing primers using NCBI primer tool, the explanation for Tm value is "The Tm calculation is controlled by Table of thermodynamic parameters and Salt correction formula (under...
02 February 2016 5,657 11 View
I'm trying to amplify a 8kb fragment (gene+promoter) form genomic DNA, using roche Expand™ Long Template PCR System. My primers are ~21bp long and the annealing temperature is 2 degree centigrade...
01 January 2016 8,779 11 View
