What the order of the substance to be added for restriction enzyme digestion?
Water
10x NEB Buffer
100x BSA (opt.)
RE enzyme
DNA in ng/uL
Then when we should mix all this component? Is it after RE be added or prior RE is added?
Hi there,
Water first then buffer bsa and DNA. I always add RE the last and homogenize the complete reaction mixture using pipetman or giving a flick to the tube and then centrifuge to put the liquid at the bottom.
Thank you so much
I stored my buffy coat sample at -20C. So,what is the best n proper way to thaw the sample?vortex?centrifuge?duration?
03 April 2017 9,258 0 View
I want to ask concerning on QuantaSoft software. For copy number variation (CNV) what the type i have to choose for FAM as well for VIC.
02 March 2017 10,076 0 View
How long to thaw the component for TaqMan Copy Number Assay? What is the rpm,degree Celsius, duration to centrifuge the component for TaqMan Copy Number Assay?
02 March 2017 3,890 4 View
Is it a necessary buffy coat I aliquots for extraction of DNA is added with RNA later prior to storage at -80C?
01 February 2017 1,189 4 View
So that i can assemble directly in ddPCR SuperMix
01 February 2017 5,647 2 View
How many times i can freeze and thaw? -For the component as below How long to thaw? -For the component as belowOnce i thawed can i stored back at -20C?-For the component as belowHow I should...
01 February 2017 5,753 2 View
How many times i can freeze and thaw? -For the component as below How long to thaw? -For the component as below Once i thawed can i stored back at -20C?-For the component as below How I should...
01 February 2017 2,289 23 View
Is it necessary to use MULTI_CORE 10X Buffer fro digestion of DNA?
01 February 2017 9,852 1 View
01 February 2017 4,490 5 View
How long can I stored the digested DNA at -200C ? And Then if I dilute the DNA digest, can i store at -200C? How long i can stored it(duration)? OR Is it better not to dilute the DNA and stored...
31 December 2016 3,088 6 View
Does anyone tried to do nucleofection with AMAXA by Lonza with lower than recommended amount of buffer in the cuvettes (100 ul)?
07 August 2024 4,616 0 View
Currently, when I run SDS-PAGE, I don't see any bands at all, even though I used the same material just a day ago and it worked fine.... In our lab, we dilute the 10X running buffer to 1X and...
06 August 2024 5,373 2 View
I want to introduce a point mutation (change in one nucleotide) into my gene of interest (DNA binding domain) I have designed primers as recommended on the Data sheet of the kit : -Both primers...
05 August 2024 9,059 3 View
Hello everyone, I'm encountering an issue with my electrochemical impedance spectroscopy (EIS) measurements and would appreciate some insights. Experimental Setup: Electrodes: Gold interdigitated...
05 August 2024 3,783 2 View
I'm interested in learning about enzyme usage. Could someone explain the basics of working with enzymes? I understand they're different from standard chemicals, but I'm uncertain about the...
01 August 2024 5,460 5 View
Is it the "elution buffer" or the "dialysis buffer"? Note: I'll be using NanoDrop OneC
01 August 2024 967 3 View
I am trying to adapt adherent CHO-K1 cells in F12K+10%FBS to suspension culture in only F12K, but in vain. I have followed Mirus bio as well as Thermo protocols, please suggest any alternative....
31 July 2024 3,756 0 View
Recently I'm trying to use Sep-pack C18 1 cc Vac Cartridge (Waters: SKU: WAT054955) to desalt digested peptides. However, the recovery rate is usually 40-60%. What is the normal recovery rate?
31 July 2024 6,544 11 View
I want to choose a resarch topic regarding enzyme inhibition. So I did my research and found out most of the diseases that originate from enzymes were actually caused by the "deficiency" of...
30 July 2024 2,483 5 View
We have encountered an issue with the serum of mice being hemolyzed after the process of centrifugation. Could you please provide guidance to prevent hemolysis ?
29 July 2024 8,402 4 View