What does XM_xxxxxxxxxx mean in NCBI? - FAQS.TIPS

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Can anti-6xHis antibody detect 8xHisTag?

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...

10 November 2017 6,509 5 View

Anyone use this PCR mycoplasma detection protocol and get a 100bp product?

Hi researchers, I'm using a PCR mycoplasma detection protocol which is found online and it works well for me. Link as...

09 October 2017 4,399 4 View

What is the difference between I-TASSER and QUARK protein structure prediction?

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...

07 August 2017 2,305 2 View

Positive control gene for real time PCR in IL6 stimulated cells?

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...

06 July 2017 6,054 3 View

Why cannot detect protein after over-expressing by lenti-virus?

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...

06 July 2017 1,741 4 View

Is Swiss prot ID unique for each transcript?

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...

03 April 2017 2,391 3 View

Secretory protein with Flag at N-terminal?

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...

31 December 2016 6,995 7 View

What is the size for BMP2 protein in western blot?

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...

31 December 2016 7,419 4 View

Which part of the siRNA matters most?

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,...

09 October 2016 499 4 View

For secreted protein, can we do western blot using cell lysiate instead of ELISA of cell culture supernatant?

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...

08 September 2016 1,781 4 View

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Can I base on reverse DNA sequences to perform alignment, convert to amino acids and GenBank submission?

I have reverse sequences (AB1 format), can I base on reverse DNA sequences to perform nucleotide alignment, convert nucleotides to amino acids and deposit the sequence in GenBank database?

11 August 2024 5,138 1 View

How to confirm the site-directed mutagenesis result without performing NGS?

I'm cloning a fragment of 3200 nts into plasmid. The cloning was successful, however, 02 amino acids were mutated. Now I want to fix these 02 aa by site-directed mutagenesis technique using...

08 August 2024 4,645 2 View

I can't see the ssDNA band after performing asymmetric PCR. Is there any way to do this?

After performing symmetric PCR, PCR purification was performed. Afterwards, asymmetric PCR was performed using the PCR purification product as a template, but no ssDNA band was confirmed in the...

08 August 2024 1,668 3 View

Why after performing site directed mutagenesis ,I don't see any colony after transformation?

I want to introduce a point mutation (change in one nucleotide) into my gene of interest (DNA binding domain) I have designed primers as recommended on the Data sheet of the kit : -Both primers...

05 August 2024 9,059 3 View

Anyone having idea about VN primer for miRNA primer design ?

How to design VN primer to attach with universal reverse primer

05 August 2024 2,116 3 View

Why did the authors extrapolate a phenotype that they experimentally proved in one bacterial strain across the whole genus of the organism?

I aim to be as skeptical as possible regarding whether a pair of orthologous genes results in the same phenotype in their different but related bacterial organisms under similar environmental...

05 August 2024 6,787 4 View

Why my colony PCR results of my recombinant bacterial not showing any results?

I am performing ligation of the plasmid and a target gene. The steps I have taken are: 1. Double digestion of the plasmid and target gene 2. Ligation of the plasmid with the target gene 3....

05 August 2024 2,570 3 View

Does anyone have issues using Prepman Ultra reagent for MicroSeq ID bacterial, fungal and yeast sample preparation?

I have been attempting to extract DNA from Bacterial, Fungal and Yeast banked samples (>1e7 cells) using Prepman Ultra reagent and I seem to be struggling to obtain a sequence. Although the...

01 August 2024 2,079 0 View

I'm optimizing a tetra-ARMS PCR protocol with amplicon sizes: 165bp, 123bp and 93bp. Why, in the gel, only 165bp is visible while I'm expecting all 3?

It's an end-point PCR protocol. I'm using 1.5% agarose gel with SyBR Safe dye and TBE as a running buffer, visualization on BioRad XR+ system. I was primarily thinking of primer efficiency,...

01 August 2024 4,673 4 View

What is the acceptable p-value cutoff for GO enrichment analysis ?

I have an RNA-seq data that I have analysed using Limma-voom and have extracted the gene IDs, log2FC and the p-values. At p value < 0.05, I have over 10,000 DEGs, however, when I run the GO...

31 July 2024 225 2 View