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Questions related from Soundarya Priya
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...
08 August 2019 3,658 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
05 May 2019 2,182 3 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...
02 February 2019 2,142 13 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...
02 February 2019 2,062 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??
02 February 2019 9,832 6 View
The crucial points that need to be considered to avoid failures in cloning.
01 January 2019 10,074 2 View
What is the acceptable range when checking the primers using Primer-BLAST
01 January 2019 7,209 4 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?
01 January 2019 2,736 6 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...
01 January 2019 5,541 9 View
I face many difficulties in gel purifying the restricted products A260/280 ratio is high sometimes Failure to recover DNA from gel ( completely) Loss of DNA - lesser yield So is there any other...
12 December 2018 8,960 22 View
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...
12 December 2018 508 0 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...
12 December 2018 9,461 8 View
What happens if you perform restriction digestion with an enzyme that does not have star activity but leave them for more than two days? will the overhangs be removed or chewed off? will Ligation...
12 December 2018 8,433 5 View
I restricted digested 1microgram of my PCR product with two enzymes NcoI and XhoI. I incubated one vial overnight and another for 3 hours. The vial which was incubated overnight has got a crisp...
12 December 2018 3,837 1 View
