Bio-Rad IQ5 Calibration

I would like to do PCR array on IQ5 instrument. This machine has previously done calibration files. Do I need to do calibration again?

07 August 2017 5,564 1 View

Statistical Analysis in Western Blot

May I know which statistical analysis is suitable in western blot for 3 biological replicates? Using arithmetic means or can I use geometric mean?

06 July 2017 4,005 0 View

Protein Pellet Resuspension with how much DTT?

I have extracted my protein using isopropanol from Tri-reagent LS extracted cell culture. I found the pellet is difficult to dissolve in 1% SDS. I have read through some articles saying that...

31 December 2016 2,722 3 View

How can I analyse gene expression time course study by qPCR?

I am doing time course study for gene expression in cell culture. I am doing 0, 1, 2, 4, 8, 16, 24 hours. Each time point I have treated and untreated (control). I should normalised my individual...

11 December 2015 4,659 7 View

Do gene expression real time RT-PCR study have a big biological replicate differences?

I am doing real time RT-PCR of human cell line. May I know if the gene expression in different biological replicates will be different or almost the same?

10 November 2015 9,494 3 View

How do I perform a SPSS one way ANOVA test with post hoc test significant value?

I am doing on cell culture and I have done 3 biological replicates (n=3) to get my data. I treated my cells with cytokines with five different doses and is compared to control (untreated). I get...

04 May 2015 6,271 9 View

How can I combine 3 biological replicates qPCR result (in 3 diferent plates) using CFX manager Gene Study?

I have done my 3 plates for 3 biological replicates for my control and the samples. May I know how to combine all these 3 results into one? I know this can be done through CFX manager Gene Study...

04 May 2015 6,651 2 View

How can I narrow broad melt peak in qRT-PCR?

I am doing 1 step qRT-PCR using quantifast SYBR RT-PCR and using the recommended protocol by Qiagen. I get pretty narrow peak for positive control (HepG2) but not really for THP-1 induced...

01 February 2015 4,493 2 View

Any advice on Primer Design?

I have designed a set of primers with the amplicon length of 137bp, I have done my qRT-PCR but it is not specific . I have done the sequencing but the result is only showing the 23bp which is the...

11 December 2014 6,093 10 View

Can I optimize dose and time in different cell lines?

I am doing dose and time response in animal cell culture. Do I have to optimize the dose and time in one particular cell line first, then use the optimized dose for another 2 cell lines? Or, do I...

10 November 2014 5,532 2 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

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

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

RNA-seq DEGs downstream analysis and reporting in literature?

I used EdgeR to get the list of DEGs (differentially expressed genes) from my liver RNA-seq experiment. I have done the gene overrepresentation and GSEA based pathway analysis. I am currently...

23 February 2021 8,969 1 View

Could you please suggests a tutorial or paper or any other materials to learn multiple gene expression in TCGA and GTEx databases?

I would like to analyze multiple gene expressions in normal and cancer tissue by using the data in the TCGA and GTEx databases. Please suggest a tutorial or paper showing step by step analysis of...

23 February 2021 9,007 3 View

Are there any databases for gene co-expression or expression pattern clustering?

I am currently working on gene clustering based on co-expression pattern in mouse brain. The problem is I do not have some solid way to test my result. Are there any suggestions for databases...

22 February 2021 4,374 3 View

Which CpGs have a significant effect on gene expression?

It is known that methylated CpG around TSS decreases gene expression. However, It is difficult to know exactly which CpG has a greater impact on gene expression. Are there any databases known...

20 February 2021 5,634 3 View

I can detect my protein but the encoding DNA sequence has stop codons within. Should I ignore the stop codons?

I constructed a synthetic DNA library (scFv, VH-VL orientation) with a 3' reporter and 8x histag. I cloned and expressed this gene following which I performed an ELISA. The ELISA results suggested...

19 February 2021 7,006 5 View

Purification of proteins from baculovirus-infected insect cells?

18 February 2021 6,665 6 View

Found that there was a multiple band when amplifiying targeted gene ?

i few days ago amplified specific gene using PCR with specific primers, but there was a multiple band in electrophoregram i found. out of 10 sample amplified, there was multiple band and there was...

18 February 2021 5,131 5 View