How many nucleotides are required on both sides of restriction site by the enzymes like NdeI and BamHI for restriction?
Usually 3 nucleotides is OK for most REs. You can check this on the web site of the suppliers (New England Biolabs, ThermoFisher)
Dear Muhammad
you can find the flanking sequences suggested from NEB for each enzime at
https://international.neb.com/-/media/nebus/files/chart-image/cleavage_olignucleotides_old.pdf?la=en&rev=c2f94e1cdcd549c5bf8fdb59f7b63f67&hash=4FBF94A3878D7C5E79BC1DBBADD5CCDF70803D06
Ciao
Manuele
Manuele Martinelli, many thanks for sharing the useful page. We have added 8 to 11 bp at the 5' end of our gene sequence and initially it has given better results.
I have dataset which shows the length of power lines. I need to classify the lines based on the line length. Is there a rule to classify the High voltage (HV) and low voltage (LV) lines based on...
03 March 2021 4,116 4 View
Actually, I am running cfd simulation for a heat exchanger which has two fluids one hot and other one is cold fluid. there is a solid domain between them, which I removed and instead I have used...
01 March 2021 9,537 2 View
I am simulating Heat Exchanger. I have tried and tested all the methods to resolve this issue from the internet like refining mesh, improving skewness and orthogonal quality ,extending outlet to...
01 March 2021 6,985 3 View
I have to amplify a gene and my primers just reached. The Tm for Forward primer is 64.2, and that of reverse primer is 65.5. Can some one suggest how to get the best annealing temperature? Thanks...
01 March 2021 360 7 View
Should I mesh the solid domain ? or should I use wall thickness? or Shell conduction? for Heat Transfer between two fluids of a Heat Exchanger. what is recommended?
01 March 2021 9,785 3 View
I have a virtual machine. My host Pc specs are not that good thats why i went for virtual servers. so basically my host pc has 8gb of ram with nvidia quadro k2200 4Gb DDR5 GPU ,processor intel...
01 March 2021 8,415 1 View
Is the period to autoclave not enough? The inoculation in a hood and flame and UV and alcohol.
27 February 2021 9,356 3 View
hello. i am junior researcher. I received a comment from a reviewer that he could not find NMR data in my submitted paper. I had included the NMR graph with explanation and formula to calculate...
27 February 2021 734 4 View
I have a sample of treated water sample & asked to obtain ammonia conc. in sample using a machine with 5-25 ppm detection range. However, upon testing with different dilution factors, all...
27 February 2021 8,483 3 View
I am working on TiO2 composite material and the results of XRD have a lot of small/junk peaks and while I apply a smooth filter in Origin then the other characteristics peaks disappear so anyone...
26 February 2021 8,183 3 View
I am trying to identify these 3 genes among some tomato cultivar collections and after aligning some sequences from NCBI, I couldn't find unique sequences to target for specific primers. There...
28 February 2021 606 3 View
I have cloned two domains of a gene into two bacterial vectors using cut and paste cloning with different RE sites. I checked the putative clones by colony PCR and Restriction Digestion (fallouts...
18 February 2021 6,736 4 View
Hello, I have designed a specific primer for my Gene, when I blast this primer, it just finds the mRNA of the gene, but in nr blast, I have found some CDSs that are not relevant to my Gene. Do...
05 February 2021 5,842 3 View
Hi everyone, I need to clone in pGEM-T vector the result of amplifying a metagenomic library by PCR with a high fidelity polymerase but I only want to clone the fragments of an specific size...
04 February 2021 6,051 8 View
03 February 2021 221 2 View
01 February 2021 9,576 3 View
Due to the pandemic of coronavirus, the most source that the people depend on was social media. There is no guaranteed solution that is biased from political opinion. An example of that the...
01 February 2021 7,567 3 View
The recommended amount of DNA for restriction digestion is 1ug in a 50ul rx I've got around 500-600ng of DNA in a total volume of 35ul. Would I get satisfactory results with this amount of DNA for...
31 January 2021 729 6 View
Hi everyone, I perform a PCR + Restriction Digestion (single cut, same at both sides) over a metagenomic library. After that, I am trying to purifiy DNA fragments between 1-4kbp long from the...
24 January 2021 2,190 5 View
Considering that we hypothetically designed an experiment, we have an x gene, and we have the mutant version of this gene with a single nucleotide mutation in one of the exons. What kind of an...
22 January 2021 9,589 12 View