21 Questions 2 Answers 0 Followers
Questions related from Alexander Supertramp
I would like to colour a protein structure in heatmap-like colour scheme. I have a list of values for each residue and would like to represent on the protein structure (eg. more positive = blue,...
30 August 2021 7,841 3 View
Zymo sell kits for yeast plasmid isolation, however I have read that you can basically just digest the yeast cell wall separately (with zymolyase treatment), then just use a Qiagen miniprep (for...
29 April 2021 1,890 3 View
Sequence diversity is important for good reads on illumina seq platforms, particularly at the beginning, however is it that important AFTER the clusters/ colour matrices have been identified/...
01 March 2021 8,245 1 View
I transformed a plasmid into yeast and selected for transformants by growing them in a selective media (liquid, not agar plate). My experiment requires that I continually grow/ split these...
02 February 2021 10,090 2 View
I typically don't adjust the pH of my CSM (SC-HIS) or YPD media when growing yeast, and wondered if it had a significant effect on growth? After measuring both, YPD seems fairly neutral however...
26 November 2020 1,931 6 View
I am also a little confused as to what happens if you were to take mid-log cells (for examples' sake say OD600 ~0.3-0.8) and dilute to below 0.3. Do they slow their growth rate in a quorum...
13 November 2020 6,467 1 View
I am having difficulty finding an answer to this question - what is the maximum DNA insert length feasibly permitted by MiSeq. By 'DNA insert', I mean EXCLUDING the adapters. And does anyone know...
05 November 2020 4,882 3 View
I can find a lot of information on what concentration you use to select for G418-resistance s. ceverisiae, however I can't find much on the concentration at which even resistant cells die. For...
02 September 2020 9,242 3 View
I am currently trying to generate yeast growth curves in a less labour-intensive way using a plate reader that has the ability to shake and incubate. Currently I have trialed a Tecan Infinte Pro...
22 July 2019 5,042 2 View
I've purified my plasmid from DH5a for a double digestion (NcoI and XhoI), however I've noticed it only appears to be cleaving at one site (ie I'm only getting one band in the gel). Previously it...
07 July 2019 1,966 1 View
I am still relatively new to working with yeast, so was wondering what happens if you leave yeast at 30 for longer than you need to so that they go on to form big fat colonies? I imagine on...
06 June 2019 1,719 6 View
I have come across some protocols that suggest using pre-cultures that are cells grown from a plate colony over the weekend to stationary then left on the bench at room temperature, that you can...
06 June 2019 6,299 1 View
06 June 2019 1,227 1 View
I have pre-made LB and YPD bottles of agar for pouring plates in autoclaved duran bottles - how long are these good for? Do they begin to degrade under these conditions?
09 May 2019 1,759 3 View
05 May 2019 2,066 3 View
I'm trying to find the minimum inhibitory concentration at which a series of yeast strains grow in the presence of the antibiotic Aureobasidin A. When I plate the yeast on agar they show a trend...
20 February 2019 5,190 4 View
I have previously confirmed the presence of my (desired) plasmid after transforming it into bacteria and sequencing a colony. If I then transform that (purified) desired plasmid again into...
08 January 2019 7,321 8 View
I have an DNA insert that I intend to order as a complete double stranded construct and clone into a vector using restriction enzymes, however I am unsure exactly how to design the restriction...
01 January 2019 9,901 3 View
Can anyone point me in the direction of some literature/ give me advice on how to design linkers between fusion proteins? I appreciate there's probably no single perfect way to make them, but as I...
01 January 2019 7,445 4 View
01 January 2019 7,561 7 View
I am trying to obtain a stronger readout from an experiment that involves a transcription factor binding a target DNA sequence. I have experimented with single and triple tandem copies and there...
01 January 1970 2,325 1 View
