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Questions related to Molecular Biological Techniques
Can anyone suggest how to decide the annealing temperature from the Tm value of primers?
08 September 2016 7,680 4 View
Dear friend, I am trying to use yeast strain BJ2168 to over-express Gal1 promoter driven GFP. I grow the strain in SC complete media-glucose to OD 0.5. After washing the cell with PBS, I replace...
08 September 2016 9,745 2 View
Do you guys know a cheaper alternative to sybersafe for running a gel?
08 September 2016 9,853 14 View
Hi, Anybody knows how can we quantify total amount of specific protein i.e X protein from western blot of cytosolic and nuclear fractions? Thanks in advance. Best.
08 September 2016 2,083 3 View
I need to preserve DAN RNA samples for shipping via travel, for a period of 6 to 12 hours. How to storage and transport them in RT or other packaging methods ? and what is the protocl of freeze...
07 September 2016 9,010 4 View
Hi, I would like to estimate the best time point to observe IFN-g mRNA in T cells activated with aCD3 stimulation. In this condition, I have already verified IFN-g protein production at 48h.
07 September 2016 7,558 2 View
I am working on a virus that we have in a bluescript vector. The end goal is to rtPCR patient data but to test the primers I'd like to use our viral genome without blue as some of the primers will...
05 September 2016 5,413 4 View
We just bought the enzymes to investigate the polymorphisms in H pylori biopsies. When the student was preparing the G buffer to add in the restriction enzyme tube she added TAE instead ultra pure...
05 September 2016 5,043 3 View
If anyone has done GCMS for tetrodotoxin detection, please help me to know the compound names for the characteristic fragments of 2-amino 6-hydroxymethyl 8 hydroxyquinazoline that forms peaks at...
03 September 2016 3,352 2 View
What will be the percentage of my agarose gel , while running V3 region of 16SrDNA gene ?
03 September 2016 7,602 2 View
Suppose if mRNA is showing up regulation of a gene through qRT-PCR, then, is it necessary that the level of corresponding protein will also be high in western blot.
02 September 2016 1,547 4 View
First, I optimized the primers which were designed for specific exons, with different Temp (annealing Temp. of PCR) in gradient PCR with standard human blood DNA. After getting annealing Temp. did...
02 September 2016 1,268 5 View
My plasmid was purified using a mini prep and was eluted in EB. I sending the plasmid off for sequencing for the first time and the submission guidelines suggested to submit the samples in water...
01 September 2016 1,664 2 View
I have cloned my insert DNA into pBluescript SK+ vector, after this cloning I have performed site directed mutagenesis by using PCR then digested it by DpnI digestion. what control should I use...
01 September 2016 7,777 8 View
I am trying to make calcium alginate beads for cell immobilization, sodium alginate solution was prepared in the percentage of 2, 3 and 4 with worm d. water. The sodium alginate solution was...
01 September 2016 313 3 View
Hi everyone. I'm trying to label my probes with FITC and Cy3 by PCR. I've tried several protocols (different ratios of labelled and unlabelled nucleotides) but the PCR products don't seem to be...
31 August 2016 251 1 View
I've been trying to amplify the products for quite a while now and I have difficulty in getting good bands or any bands at all, I'm wondering if it's the storage condition of the components or...
31 August 2016 1,169 7 View
Has anyone had any experience with storage tissue in RNA later for RADseq? I know that RNA later is suitable for DNA storage, but was wondering about the quality or any complications that might arise.
30 August 2016 3,344 2 View
I do not have a plasmid containing the NLS. I was planning to add the NLS to the sequence of a primer. My question is: Do I just place the sequence straight after the Start codon without a linker...
30 August 2016 874 4 View
by using the overlay method of antagonism the results are good and there is a clear zone but with ethyl acetate extract there is no results.
27 August 2016 6,480 5 View
I am totally confused about the RNA agarose gel. I have isolated total RNA from S. aureus. I want to see it in agarose gel. I ran 1% gel with formaldehyde and added 1 ug/ml EtBr during casting....
26 August 2016 8,125 7 View
I am looking for a reagent that makes the lysis of blood cells in order to improve the yield of DNA recovered by the use of total DNA extraction kit. Does anybody know this reagent ?
25 August 2016 2,847 1 View
Hi, I am studying genes for the resistance to the antibiotics in Klebsiella and I am analysing an outbreak of carbapenem-resistant isolates. The gene I am analysing is OXA-48 and it is known to...
23 August 2016 934 3 View
Hi, what is the best protocol to isolate DNA from FFPE tissue. I have tried KAPA Express Kit, NucliSENS Easymag, Organic Extraction, please advice further. Thank you!
23 August 2016 7,781 6 View