How to enhance stability of c-DNA?

More Ravindra Deshpande's questions See All

Any advice on my real time PCR amplification problem?

I am having problem with real time (SYBR green) amplification of gene. I have designed primers spanning exon-intron junction. Primer con, was 1picomole. I have used 5micrograms of RNA for cDNA...

02 March 2015 3,971 12 View

What are the best kits for miRNA isolation?

I am planning to isolate some miRNAs fro human brain tissues as well as from cell lines. Can anyone please suggest kits?

03 April 2013 6,364 4 View

Mirna prediction tools

Can anyone suggest a computational tool or software to predict mirna- mrna binding (3', 5' UTR and in ORF too.)?

03 April 2013 3,029 6 View

Primer designing for miRNA.

I am planning to detect expression of miRNAs in human tissues. Could anyone please let me know how to design primers/any online resources to design primers for miRNAs? Is it similar as we design...

03 April 2013 152 11 View

miRNA - mRNA interaction?

I want to check whether some specific miRNAs have putative binding sites in genes specifically participating in apoptosis process. Can anyone please suggest me some computational tools for the same?

03 April 2013 7,152 2 View

What is a good software to predict protein-protein or protein DNA interactions?

Can anyone please suggest tools or software for predicting putative protein-protein or protein-DNA interactions?

11 December 2012 9,647 6 View

Gene inhibition

Can anyone suggest what could be better option to inhibit specific gene.. Sirna or specific inhibitor compounds??

07 August 2012 3,784 2 View

Virtual PCR

Can anyone suggest any online bioinformatics tools to check theoretical amplification for primers I selected for a specific sequence?

07 August 2012 8,001 10 View

Software to predict putative DNA or protein binding sequences

I would like to check whether my protein of interest has putative DNA or protein binding sequences in it. Can anyone please let me know bioinformatics software and tools available for predicting...

06 July 2012 5,642 0 View

Gel electrophoresis result for total RNA samples, 1.5 Agarose gel was used, How to improve the integrity of RNA?

Gel electrophoresis, RNA degradation, RNA extraction from fresh tissue

02 March 2021 5,433 5 View

Expression cloning by using cDNA,do I need to modify the cDNA first?

I am going to have a expression cloning of mammalian gene by using shuttle plasmid to transforming the E.coli However I don't know I should only inserting the Coding sequence ,or I can...

28 February 2021 5,440 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

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

Why is the efficiency of viral mRNA electroporation in PBMC low?

Hello, I have been struggling with the electroporation of PBMC with HIV-I mRNA. Although the quality of the RNA generated is good, and the handling of the cells in meticulous, the frequency of...

27 February 2021 9,233 3 View

Why do the PCR bands from the cDNA templates (extraction from the cell after transfection) show different size from the band of the plasmid template?

After transfection with the plasmid ( linearized ) and subcloning of the cell lines, RNA was extracted from the cell and then reverse transcripted to cDNA. When PCR reactions were run to verify...

25 February 2021 5,712 3 View

What is the best concentration of cDNA to be used for qPCR?

I have synthesised cDNA from 500ng/ul of RNA. The concentration of the cDNA Is about 5000ng/ul. Should I dilute my cDNA or just use it as it is for my qPCR? I usually use 2ul of cDNA for a 20ul...

24 February 2021 8,618 3 View

Recommendations on homogenizers for RNA isolation?

I will be performing RNA isolation with frozen lung biopsy samples and am in the market for a tissue homogenizer. I am wanting a homogenizer that will fit in a 1.5ml tube. I was looking at the...

24 February 2021 3,623 3 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

Is it efficient to do liquid biopsy to investigate the gene expressions of the tumor?

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Thomas Manesh

cDNA is stable, it is not unstable like RNA. Even u can store it for more than 6 months if your cDNA is contaminant free.

Ajay Saini

Hi Ravindra

as mentioned above by Thomas, cDNA is more stable and can be stored for a long time....you can ensure that by purifying it before storage, and also by storing in aliquots (to avoid repeated freeze thawing) in case you are planning for a long term use..

bye

Ravindra Deshpande

Thank u Ajay and Thomas... what would be appropriate temperature to store cDNA and is it OK to prepare its aliquots in mili Q?