19 Questions 109 Answers 0 Followers
Questions related from Tias Saha
Bacterial cultures are generally grown at 180 rpm in a shaker incubator. Is it advisable to use it below 180 rpm? Due to shaker malfunction, I'm unable to use it at 180 and I have to shake it at...
09 August 2017 8,864 24 View
I have a WT and null cell for a particular protein. I also have complemented set of cells which can be selected using kanamycin. For experimental issues I cannot use kanamycin. Will the plasmid be...
06 July 2017 2,340 3 View
I'm trying to revive glycerol stock of E. coli DH5alpha by streaking on a Nalidixic acid (NA) containing plate. I have prepared fresh nalidixic acid (15ug/mL) but I'm getting mixed reviews on the...
07 June 2017 1,038 9 View
I'm using RPMI 1640 media from Himedia. Within 24 hours the colour of the media is changing to yellow even with very low density of cells. I have checked for any kind of contamination, but that is...
06 May 2017 3,906 13 View
I have to purify a bacterial protein from E. coli. My protein's yield is quite low. I have read that batch binding gives better yield, if so, for how much time do I have to incubate my protein...
06 February 2017 1,149 5 View
If I have a sequence of amino acids. Can its tertiary structure be predicted using any in silico tool? There are proteins which have not been crystallised yet and hence we do not know the...
09 August 2016 3,260 8 View
a. As protein folding generally starts from N terminus. If hypothetically I delete some parts of its N-terminus, will it still be stable and able to fold properly? b. If I delete 30 amino acids...
20 July 2016 357 8 View
I feel it contradictory to evolutionary advantage that the two strands of DNA as evolved are anti-parallel but the polymerases that synthesize/ replicate them can do so in only one direction....
13 July 2016 806 15 View
I need a quite a large number of cells to check the expression of certain constructs. Right now I do not have that many cells growing. Can I thaw a frozen vial and without passaging, use them for...
22 June 2016 8,116 6 View
For plasmid vector, linearization is the indication of complete digestion, but how to check whether the insert which is a linear PCR amplified fragment is completely digested?
28 May 2016 9,523 3 View
It is often recommended to heat inactivate restriction enzymes, ligase, phosphatase at 65 degree for 10 mins or so and use the DNA for subsequent steps of cloning. How efficient is the heat...
16 May 2016 7,264 4 View
I eluted my amplicon from gel and checked its concentration after elution (column based kit) on 0.8% agarose gel. In the eluted product I found another band above my amplicon, which is very...
27 April 2016 5,879 13 View
I used colony PCR to screen some colonies for positive clones using the gene specific primer. I got 9 out of 10 colonies which gave me the amplicon (a mammalian gene, unlikely to be present in...
20 April 2016 2,603 3 View
How does the reaction volume determine the efficiency of restriction digestion? I have 200ng/uL insert in 30uL of buffer. What should be the reaction volume [water, buffer, enzymes (20U), DNA...
06 April 2016 3,796 2 View
I loaded approximately 20ug protein in each lane. After electrophoresis, the outermost lanes appear to have been spread outward rather than following a linear path. What could be the reasons...
28 March 2016 9,935 21 View
In terms of loss, quality and successful cloning, which method is better to use for purifying the PCR amplified insert and the double digested insert, Phenol-Chloroform -Isoamyl alcohol extraction...
28 March 2016 2,966 7 View
I want to study truncated versions of a protein to analyse its domain specific function. So, for detection I need to put some tag at the C terminal as I have N terminal deletions. I have used HA...
03 March 2016 5,252 6 View
Are the components of DMEM media used for cell culture susceptible to UV degradation or modification? Is it advisable to keep the media under UV in the biosafety cabinet for a while? can that...
12 October 2015 4,948 4 View
Why does the freshly prepared media seems to be yellowish red, while the media bottle opened many a times turns purplish red? Even used media (yellow colour indicating acidic pH) when kept outside...
13 July 2015 8,337 1 View
