I need to get out cDNA from a plasmid which has an internal EcoRI restriction site. The weight is around 9000 pb. EcoRI is the only enzyme that could do this job.
Thanks a lot, the second option sounds really good!!!!
Low level of digestion enzyme can be used for partial digestion
ok, but how many DNA I have to put in the reaction mix... I tried 0.1 units of enzyme, with 1 ug of DNA for 30 min... and all the DNA was digested!
I've doing some recordings using clampex, the file was generated on the lab book and the files are there. However i opened them today and i realize that the recordings were not there. I have no...
10 November 2018 1,050 0 View
I would like to know if my protein of interest is glycated and if it is target of RAGE. Is there a specific experiment to know if a protein is glycated? Do you know if there is a form to know...
09 October 2014 891 10 View
Hi, I have problems with running gel electrophoresis. I have tried agarose gel electrophoresis and native PAGE. I have two proteins, which have molecular weights of ~30kDa and ~180kDa and two...
03 March 2021 4,275 4 View
Gel electrophoresis, RNA degradation, RNA extraction from fresh tissue
02 March 2021 5,433 5 View
I'd like to perform single-strand conformation polymorphism (SSCP) in my thesis, however I cannot control the temperature of the vertical PAGE since we are using the conventional tanks. Is there a...
02 March 2021 9,157 1 View
Can someone please give me some possible things that could go wrong? Here is my recipe: 0.5g Agarose 50 mL of TAE 1x 1 uL ethyl bromide. Gel was run at 100V for 1 hour. The buffer used is also TAE.
01 March 2021 9,952 3 View
Hi, I am running a size exclusion chromatography experiment with a buffer containing Potassium Acetate as a salt. I analyse these fractions through SDS-PAGE. After boiling my SEC fractions in...
01 March 2021 2,622 3 View
I am worried about this overexposure of the upper part of the gel in the picture. Is it possible that this is the result of too much concentration of ethidium bromide? Why is there such a big...
25 February 2021 8,140 3 View
I tried to separate my substance from a protein in a sillica gel column. By mistake, it dried and cracked. Can I save the trapped substance? Can I repack it ,or at least elute all of the trapped...
22 February 2021 1,789 3 View
This is gel electrophoresis result for total RNA samples with good conc. However, I guess there is a problem with the purity of the samples, i can't figure out why it appears like this? And how...
22 February 2021 338 9 View
Hi all, I proceed with nucleic acid gel electrophoresis recently several times, but a wried phenomenon is they always melted after running over 30min. My gel concentration is 1% percentage...
22 February 2021 9,896 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