Tatpong Boontawon

13 Questions 18 Answers 0 Followers

Questions related from Tatpong Boontawon

If I coat protein for indirect ELISA overnight , Should I add PMSF into coating buffer or not?

I express recombinant protein in yeast. After that harvest cell and resuspend cell in Phosphate Buffer Saline (PBS) buffer with 1 mM PMSF (as protease inhibitor), then break it by glass bead and...

29 September 2016 3,546 3 View

Does anyone have an experience with sucrose cushion centrifugation for virus?

I try to purify virus-like particle that express in recombinant yeast. By sucrose cushion, then subject to equlibirum density CsCl centrifugation. After I culture yeast for 100mL, harvest, break...

21 April 2016 1,193 3 View

Does ethanol denaturing enzyme, and causes non-function of protein?

I use Pichia pastoris for recombinant Lipase enzyme from Candida sp. production (secreted into medium). I try to follow the graduated members that do this project, unfortunately the protocol not...

05 March 2016 8,701 4 View

Does anyone use re-boiled Salmon Sperm DNA for Yeast LiAc transformation and get a low efficiency?

I use Lithium acetate transformation method for transforming intact circular plasmid into yeast. In past, I always aliquot a new salmon sperm DNA from commercial tube, then boil in water for 10...

26 August 2015 1,786 8 View

Could anyone suggest the Deletion of which gene can reduce hyperglycosylation in Pichia pastoris?

In yeast, we face to the problem of recombinant protein expression because of its ability in hyperglycosylation. If I aim to reduce the hyperglycosylation without affect to the growth of the Yeast...

25 June 2015 3,578 3 View

Can S. cerevisiae URA3 gene can function in H. polymorpha NCYC495?

I constuct plasmid that contains FMD promoter, MOX terminator, HARS1, and URA3 gene from S. cerevisiae, then transform by LiAc method to H. polymorpha NCYC495 for three times. But every time don't...

26 May 2015 8,017 3 View

Can intact circular plasmid integrates into Genome of Yeast S. cereivisiae?

I want to design an experiment to confirm that my S. cerevisiae URA3 gene can function in S. cerevisiae. My plasmid that is a BluescriptiiSK+ harboring S. cerevisiae URA3 gene (and don't has any...

23 May 2015 3,718 6 View

Does anyone suggest pBluscript II SK(+) or related with Kanamycin resistance marker

Does anyone suggest a plasmid that be a "pBluescript II SK(+) with Kanamycin resistance marker (instead of AmpR)" or relates. Best regards, Tatpong

03 December 2014 8,747 3 View

Where can I get L1 HPV52 and L1 HPV58 antibody?

I'm doing my M.Sc. project in the title "Expression of L1 HPV52 and L1 HPV58 protein by yeast" but I can't find any antibody use for detection these proteins. Can anyone tell me. How can I find...

18 November 2014 6,918 3 View

Has anyone used H. polymorpha NCYC495 for recombinant protein expression?

I try to find yeast H. polymorpha that have auxotrophic markers in many collections for use with my M.Sc. project, but I found only H. polymorpha NCYC495 ura3- or leu2- available. I read many...

11 November 2014 5,015 3 View

Could you suggest high fidelty DNA polymerase for cloning about 300-600 bp DNA fragments?

I want to clone about 300-600 bp DNA fragment from yeast genome. Since there have many high-fidelity DNA polymerase i.e. Phusion, KOD HiFi, Q5 etc. Colud you suggest me which is the most proper...

27 October 2014 6,632 7 View

Should I clone these genes or synthesize them because I have less time?

I do a M.Sc. thesis about expression of recombinant protein in Hansenula polymorpha (syn. Pichia angusta). Since there is no commercial vector available, we have to construct it ourselves. I have...

04 September 2014 7,149 8 View

Could you help me to find the codon usage table of Hansenula polymorpha?

Now, I'm doing my thesis about expression of recombinant protein in Hansenula polymorpha.Then, I have to do codon optimization of a gene in H. polymorpha, but I cannot find codon usage table of H....

09 July 2014 177 6 View