How long I can store DNA that has been digested with restriction enzymes, gel purified and eluted in elution buffer? Will the overhangs be stable (since ss DNA are unstable) after freezing in -20 for long time? Can I use them for ligation??
I am using E.coli BL21 Plyss for a recombinant protein production. The cells debris becomes too sticky after lysis. I spin down the pellet at 13000 g (that is the maximum available here). The...
07 August 2019 3,528 7 View
The acid base titration standards are working fine. I just want to know if there are any protein-ligand standards. Thanks in advance
04 May 2019 2,094 3 View
I use PCR cleanup kit from Thermoscientific. The extraction of restricted plasmid from gel failed twice but the extraction of insert which was run parallely gave me DNA with good concentration and...
01 February 2019 1,982 18 View
Can I connect the column to a peristaltic pump operated at a low flow rate and leave the other end open to elute the contaminants out of the column? Or will it disrupt the column packing??
01 February 2019 9,756 6 View
I need a huge concentration of restricted product. So can i just digest them in a 50 uL reaction with high volume of enzyme (say 5-7uL). This is just to run the digested product in a single gel...
01 February 2019 2,068 13 View
The crucial points that need to be considered to avoid failures in cloning.
31 December 2018 9,980 2 View
What is the acceptable range when checking the primers using Primer-BLAST
31 December 2018 6,994 4 View
The HiPrep 16/60 Sephacryl S-100 column that we use is clogged with some dark coloured particles at the very top. I tried cleaning up the column with 0.5M NaOH followed by distilled water. This...
31 December 2018 5,444 9 View
Should I add oligos based on the chart flanking both the ends of the restriction site or adding oligos to the end of DNA is enough?
31 December 2018 2,665 6 View
I have purified my PCR product with PCR purification kit by thermo scientific. When analysing the sample in nanodrop after purification I get a very high absorbance at 230 nm. Will this interfere...
11 December 2018 9,371 8 View
I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance
03 March 2021 3,568 1 View
Hello, If i am doing a buffer exchange for an antibody of 1mg/mL, does the elution lose protein in the process of buffer exchange? For example, if i flow through 500 uL of 1mg/mL sample, and...
03 March 2021 6,299 3 View
Is it possible to induce site-directed substitution mutation by quick-change method on linear dsDNA? or it has to be cloned in some vector? If yes, should it be treated with the Dpn1 enzyme...
03 March 2021 401 4 View
Gel electrophoresis, RNA degradation, RNA extraction from fresh tissue
02 March 2021 5,433 5 View
Question to you and THEM, the New Journal, "Integrative Psychological and Behavioral Science" -- do you not know, and have you not seen, this done before? There appears to be a core problem for...
02 March 2021 3,024 2 View
Hi, could anyone recommend a plasmid and/or protocol for reporter gene assay in S. cerevisiae? I want to assess the effect of growth conditions on a transcription factor, so I want to clone it`s...
01 March 2021 210 1 View
I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...
01 March 2021 8,544 1 View
We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...
01 March 2021 3,355 3 View
I am going to have a expression cloning of mammalian gene by using shuttle plasmid to transforming the E.coli However I don't know I should only inserting the Coding sequence ,or I can...
28 February 2021 5,440 3 View
Anyone know the amount of protein before enzymatic digestion for phosphopeptides enrichment of serume sample in Ti-IMAC? Has anyone ever use Ti-IMAC(the material is from Chinese company named...
28 February 2021 7,310 3 View