Tatpong Boontawon

14 Questions 19 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...

09 September 2016 6,995 3 View

Why In Vitro (Cell-free) recombinant protein synthesisis not used instead of microorganisms in the industrial scale?

I'm a student that do a project by using yeast for recombinant protein production, and I'm out of this field for cell-free protein production, but I have some doubt. I see many kits from many...

12 December 2015 7,895 9 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...

06 June 2015 7,459 4 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...

05 May 2015 193 3 View

What is the meaning of Em on E.coli-Yeast shuttle vector?

I read a paper Highly-efficient electrotransformation of the yeast Hansenula polymorpha (Faber, 1994) then I doubt that what is the meaning of Em or something on the plasmid? For me I think maybe...

05 May 2015 8,937 2 View

Could anyone suggest why I get very little amount of plasmid DNA after extraction from E. coli.?

I already obtained two plasmid that harboring gene synthesis A or B. First is pUC57 harboring gene A (size of insert 1.5 kb). Second is pUC57 harboring gene B (size of insert 1.6 kb). Then, I...

04 April 2015 7,888 14 View

Does anyone ever use PDM (Plasmid DNA Medium) for E.coli plasmid preparation?

I read from Bitesize Bio website that Michael K. Danquaha, and Gareth M. Fordea (2007) published a paper for plasmid DNA production. This PDM can give a higher yield than the other E. coli medium,...

03 March 2015 4,883 3 View

What is the chemical for washing and removing an undesired or contaminated Plasmid DNA?

Now, I'm in the step of plasmid construction in E. coli. But I found a big problem because of an undesired or contaminated plasmid DNA (some one called Ghost plasmid) in my transformants (more...

02 February 2015 9,038 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

12 December 2014 1,602 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...

11 November 2014 3,856 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...

10 October 2014 3,583 8 View

Should I put only primer or primer with restriction enzyme sites into Oligoanalyzer?

I'm in the step of primer design for cloning gene from genome of yeast Hansenula polymorpha. Because of my desired gene sequence in Genbank that provide only exactly sequence and don't have any...

10 October 2014 647 11 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...

09 September 2014 7,121 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....

07 July 2014 6,164 7 View