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Questions related from Goutham Nageshwar
Assuming I would like to use some chemical mediated process of precipitating RNA and polysacharides (from tissue lysate) but leave genomic DNA intact in the aqueous layer. What are the best...
07 July 2018 2,848 3 View
I have two targets A and B both are known sequences cloned into plasmids, The primers Tm for A is 54 and respective probe Tm is 63 The primers Tm for B is 59 and respective probe Tm is 69. PCR...
06 June 2018 6,327 7 View
Many commercial diagnostic kits send single reagent (Premix) to which the user must just add respective DNA and perform Real Time PCR. What are the key parameters to be considered while preparing...
06 June 2018 5,861 7 View
We have an issue with the Taqman Probe (FAM-BHQ1) based real time detection of target. What we have been observing is at higher copy numbers >2Crore we are getting very good amplification plots...
04 April 2018 7,290 6 View
Looking for midi and maxi plasmid isolation protocols from E coli (high copy number plasmid) cultures above 100ml (LB) Protocols tried so far: Sambrook's midi and maxi prep Alkaline lysis...
04 April 2018 283 4 View
I am working with development of viral diagnostic kits using Real Time PCR (Probe based) detection. As per standard procedures we do use plasmid carrying viral gene as control and to determine the...
11 November 2017 2,776 22 View
I am looking for strategies which most of the commercial companies use for production of 1 Kb DNA ladder. I did come across good journal for a 100 bp ladder where they have designed primers using...
08 August 2017 7,717 4 View
How can one calculate the copy number with following details: plasmid size 4000 bp concentration 1600 ng/ul Now I did try using thermoscientific online tool which does tell u total copy numbers...
08 August 2017 5,985 1 View
To perform a methylation specific PCR one has to treat the DNA with bisulfite. But are there any standard measurable ways to determine to how much extent the conversion of unmethylated Cytosine to...
12 December 2016 3,990 2 View
My query is I do know that we can determine the methylation status of a dna sample using Real Time PCR (Whether it is Methylated or Un methylated) but is there any method to determine...
12 December 2016 7,841 1 View
I am working with Bacillus subtilis RIk1285 (derivative of 168 W.T from TaKaRa), the maximum O.D I get when cultured in L.B broth (Flask work) is 4 O.D and in 7.5 Litre bench top fermentor...
06 June 2016 1,594 4 View
I am expressing few cellulolytic genes using few expression plasmids (Shuttle vectors). My question is if instead of maintaining your heterologous gene in plasmid, if u use an integration vector...
01 January 2016 1,675 3 View
Kindly some one let me know the amplicon size of this TA vector(control circular plasmid with out insert provided with kit ) from LIFE TECHNOLOGIES when amplified with M13 F` and R` primer . I...
06 June 2015 9,070 4 View
Consider we have any sample taken from patient. What technique can be used to identify specific miRNA present in that sample?
05 May 2015 8,530 3 View
Circulating micro RnAs are known to get translocated via Lipoproteins or through vesicular mediated transport. Did any one come across literature/personal work where micro rna's are translocated...
04 April 2015 1,355 0 View
Kindly let me know if i can autoclave trypan blue solution as I wish to add it into Agar plates to see clear zones for cellulase activity. I also have to add CMC to the agar plates, so is 1% CMC...
03 March 2015 972 4 View
Hello Fellow researchers. I want to screen approx. 10^6 colonies from a mutant library that is supposed to show cellulolytic activity. I know that congo red and CMC in agar plate will form clear...
02 February 2015 3,161 3 View
Hello I am trying to amplify a stretch of Double stranded DNA (of 1.4 Kb) using A forward primer of 112 base pairs and reverse of 21 base pairs. The Tm value of the 112 basepairs long Forward is...
01 January 2015 4,220 27 View
Any literature evidence suggesting specific genes responsible/known to trigger the differentiation of a cancer stem cell (in any cancer) back to cancer cell kindly provide the reference and...
10 October 2014 293 2 View
Differential gene/protein expression profiles on same cell line always shows varied results, why does this happen?
12 December 2013 7,756 2 View
Let us assume that I lyse some cell and now all the components are free. Now if I wish to just pull down all the DNA and RNA (nucleic acids) what type of chemistry can work? (Considering the...
01 January 1970 4,353 4 View
I have few queries wrt HLA alleles 1. Apart from homologous recombination what are the molecular consequences which lead to difference in HLA alleles. 2. What are the major differences between HLA...
01 January 1970 9,910 3 View
I wish to use carriers like glycogen, tRNA, Salmon Sperm DNA for improving the yield and purity of DNA and RNA isolations from clinical blood and FFPE tissue samples. Would like to know which...
01 January 1970 9,868 2 View
