How can I remove a non specific band at a particular molecular weight in western blotting?

The protein of my interest is of 69 k Da, but i observe a nonspecific band at 55 Kda along with the specific band, even after increasing blocking i see this band, it also stains ladder at same...

08 September 2014 9,511 4 View

Can anyone suggest best transfection method (non viral) for SCC29B (buccal mucosa cell line)?

07 August 2014 5,380 3 View

Protocol for detection of Phospho EGFR by IHC

I am standardizing IHC for pEGFR detection using vector ABC kit, and tried different retrial conditions and antibody concentrations, but I'm unable to get staining. Can anyone suggest any specific...

06 July 2013 129 2 View

Has anyone tried - AllPrep DNA/RNA/miRNA Universal Kit from QIAGEN (Cat no 80224)

This column extracts DNA and total RNA (mRNA +miRNA), so had a query that whether miRNA so obtained can be used for miRNA profiling.

03 April 2013 3,743 0 View

Which is best PCR primer for HPV (integrated or episomal) detection in oral tissues.

I use Nested PCR with MY09/11 followed by GP5+/6+ primers, but if anyone has a better set than these please let me know as prevalence of HPV in gingivobuccal cancer is very low, so I don't want to...

02 March 2013 9,480 5 View

Non specific band in PCR for HPV detection, what can be done about it?

I use PCR to detect presence of HPV in my tissue samples, In positive sample i get single specific band (150 bp) but those samples which are negative i get higher mol wt non specific multiple...

10 November 2012 5,914 29 View

Is it possible to determine the reason why one of my genes of interest is down-regulated, eg promoter methylation or something else?

In my study on human tumors, samples showed down regulation of one of my genes of interest. I am interested to know whether its because of promoter methylation or transcriptional inhibition. How...

09 October 2012 2,854 9 View

How to extract DNA and RNA from epithelium?

Normally I use QIAGEN kit for extractions from tumor tissues, but now I am planning to do it from premalignant lesions where I need only epithelial cells. I tried taking serial cryo sections and...

09 October 2012 8,145 21 View

What is the best protocol to treat culture with 5-aza-2'-deoxycytidine to block methylation?

I am planning to block methylation in my culture and check its effect on expression of particular gene, so how should i treat my culture with aza cytidine? i.e should I give continous dose or at...

09 October 2012 1,433 18 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

Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...

01 March 2021 8,544 1 View

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 3 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

Problem with shRNA transfection. Why are only two plates getting contaminated instead of all?

I transfected my LNCaP-WT cells with 3 shRNA plus their NTC two weeks ago and split two puromycin selected cell plates on Friday last week(Feb 26). I checked for GFP in the cells, and they all...

28 February 2021 4,949 3 View

Runs of nucleotides in Sanger sequencing?

I performed site directed mutagenesis, transformation, and then I sent out plasmids for Sanger sequencing and found out that there is extension of DNA just before the stop codon. I am not sure...

27 February 2021 547 3 View

Calculating volume of DNA required from DNA concentration?

Hi I am a bit confused. They are asking me to find out the volume of DNA required in ul (a total of 30-100 ng for genomic DNA) from the DNA concentration in the nanodrop reading which was 404.8...

26 February 2021 5,029 2 View

Has anyone used the Illumina ITS1 primer sets?

Hi everyone, Illumina provides a list of primers to amplify with high taxonomic coverage the ITS1 region for further fungal sequencing, but I cannot find the exact amount necessary of each...

25 February 2021 6,969 3 View

Errors in DNA sequences?

I am trying to better understand the scope of DNA replication and sequencing errors, e.g. 1. I have seen similar error rates of 10e4 to 10e5 for cell & instrumental DNA replication,...

24 February 2021 4,397 3 View

What might be the reason for obtaining two bands in plant genomic DNA extraction?

I got a thin band and a thick band i guess it would be the genomic DNA and the 260/280 ratio is 3.

21 February 2021 6,523 6 View