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 6,357 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 4,263 4 View

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

07 August 2017 2,179 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 5,939 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 1,628 4 View

Is Swiss prot ID unique for each transcript?

03 April 2017 2,260 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 6,854 7 View

What is the size for BMP2 protein in western blot?

31 December 2016 7,301 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 381 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 1,510 4 View

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance

03 March 2021 3,568 1 View

In PRIMER, is it possible to downweight certain taxa in multivariate community datasets to reflect uncertainty around identification??

I have a dataset with about 80 different species. As usual, some species are very easy to identify with certainty whereas others are more difficult, which means that I am less certain of my...

03 March 2021 8,066 4 View

PACYC PCR not working after several attempts. Would anyone have any suggestions?

So, I have been trying to run a pACYC PCR which will be used later on for a Gibson Assembly. However the PCR is not working. I have already tried gradient PCR and changing extension time; however...

02 March 2021 1,146 2 View

How to calculate the annealing temperature of a primer pair?

I have to amplify a gene and my primers just reached. The Tm for Forward primer is 64.2, and that of reverse primer is 65.5. Can some one suggest how to get the best annealing temperature? Thanks...

01 March 2021 360 7 View

Is it possible for two human neuronal cell lines to have different gene sequence for the same protein?

I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....

01 March 2021 3,607 3 View

Anybody with an idea on how do design specific primers for detecting tomato resistance genes tm-1, TM-2 and Tm2^2?

I am trying to identify these 3 genes among some tomato cultivar collections and after aligning some sequences from NCBI, I couldn't find unique sequences to target for specific primers. There...

28 February 2021 606 3 View

What is the ideal efficiency for a qPCR?

hello everyone, I need to do standard curves for my qPCR, what is the ideal efficiency range? I tried a primer (Mglu2 receptor) that gave an efficiency of 90.2%. Is it accepted?

28 February 2021 1,254 3 View

Genome-wide gene identification protocol ?

Is there any book chapter/book, webpage or research article available to understand genome-wide gene identification?

28 February 2021 8,095 1 View

MiRNA qpcr problem?

Dear All, mirna primer showing some problem in the melting curve? any idea why? As attached is the melting curve. The forward sequence is obtained from miRBase and reverse primer is universal.

28 February 2021 5,008 4 View

Any specific pipeline or parameter to analyse total transcriptomics study of brain tissue samples?

I have two groups of brain samples, control and treated for example. It was total RNA nova seq sequencing. I tried all the available pipeline like: star+rsem+deseq2, Hista+stringtie+cuffdiff,...

27 February 2021 356 6 View