Science with J

Do you need an expression plasmid that is not commercially available (e.g. a mutated variant)? Whether you want the plasmid to contain a mutated gene or a wild-type coding sequence , here is a quick step-by-step guide. First: Find the sequence The first thing is to find the coding sequence of the wild gene that expresses the protein you want. To do this, go to  https://www.ncbi.nlm.nih.gov/ and search the name of your interest gene using the Nucleotide search. Then click on the desired organism and select  mRNA . Choose the best entry for your query according to your requirement, and select the CDS  (and the protein sequence will appear on the right, a.k.a., translation ) .   Next,  copy-paste both FASTA text codes into a Word file . We will need them later. Graphic representation of a DNA molecule. Second: If you only want to express the wild-type protein... Paste the CDS directly into the corresponding ORF item from the supplier (e.g.  VectorBuilder ...

Systematic reviews:  The advice that nobody taught you 👂 If you are about to embark on the world of systematic reviews, you will probably have flooded your mind with information from courses that are not at all practical. This is because nothing can replace better learning how to conduct a systematic review than getting down to business. The PICO question If you follow the traditional method of analysis, you will have to keep trying and failing with your search algorithm , until you get a number of articles that are feasible for you or your team to analyze . To do this, start by asking yourself an unexplored PICO ( P atient/Population; I ntervention; C omparison; O utcomes)  question , and perform a search in one or more databases, using linked terms following the guidelines of boolean logic (AND, OR...) and parentheses. Artistic representation of a flowchart  (Poklepović Peričić T. and Tanveer S., 2019). But I've gotten thousands of results!😯 After your boolean sear...

You have amplified and measured the Ct of your serial cDNA dilutions... Now what? To validate your primer couple  (i.e., to measure their efficiency) , you have probably been recommended the serial dilution method. What does the serial dilution method for validating primers entail? This method involves making dilutions of your typical control sample (e.g., 1 in 4 , 1 in 16 , 1 in 64 , 1 in 256 , 1 in 1024 ) and mixing it with your typical Master mix and your favorite primer concentration.  This way you can confirm in silico whether the amplification pattern of your primers in successively diluted samples fits a trend line. DNA strand on which the PCR reaction is carried out. How to measure the efficiency of your primer couple using the serial dilution method If you measured Ct values ​​from at least 3 replicates of each dilution, calculate the Ct average of the 3 replicates for each dilution . Consider your 1 in 4  sample as a hypothetical cDNA concentration value of 10...

Hello world , 👋 I present to you this blog where you can find out about scientific tips that will allow you to shorten your learning time in the analysis of scientific data. We will also  optimize basic bioinformatics protocols and provide bibliographic searching and fi ltering guidelines that can help you in Biomedical research . In addition, we will inform you about advanced methods in the management of mathematical data related to the field of Molecular Biology , as well as image data related to the field of Cell Biology . Jaime D.L.A. Medical Writer portfolio:  https://www.clippings.me/users/jdlopezalcala Contact:  jdlopezalcala@gmail.com