Anthony Han Kang Tee

9 Questions 13 Answers 0 Followers

Questions related from Anthony Han Kang Tee

How to amplify a mixed virus population CDNA without biase amplification?

I am working to amplify a sample with coinfected similar RNA virus to check their competitive fitness. after viral rna extraction, then proceed to cdna synthesis. later i going to amplify both the...

05 May 2016 5,887 3 View

Why is there no significant CPE observed in the cells after few days post transfection, with respect to cloning?

I have transfected a clone-derived virus that contain luciferase gene as a marker within the virus genome. I have tried to transfect the virus into human muscle cells but unfortunately i found...

10 October 2015 2,043 6 View

What happened to my transfected clone-derived virus?

Hi, everyone.I am currently doing infectious clone enterovirus work. I have tried to use Mirus transfection kit to transfect my in vitro transcribed RNA into Rhabdomysarcoma cells. after 24 hours,...

06 June 2015 4,917 3 View

What happened to my electroporated infectious clone-derived virus RNA?

I am currently doing some virus infectious clones work.I have tried to in-vitro transcribe the coxsackievirus A16 (picornavirus) infectious clone to make rna, and then transfect them into HEK 293...

04 April 2015 774 9 View

Why is it my constructed antibodies don't work for virus neutralization assays?

I'm currently using two previously constructed polyclonal antibodies to test for the virus neutralization assay. Prior to the study, I tried immunofluorescence staining and both yield positive...

04 April 2015 629 7 View

Is length of poly A tails does a matter?

I am doing a work constructing molecular clone (infectious clone) of virus and i wonder whether the length of the poly A tails will affects the replication of virus? i made it one with about 48...

02 February 2015 1,054 4 View

Will too much template DNAs in in-vitro transcription affect the reaction?

As the title suggests, I am using Ribomax T7 rna production system (promega), the protocol shows that 100ul reaction requires 5-10ug template dna. but i mistakenly put in about 4ug of template DNA...

01 January 2015 2,815 1 View

Why is my plasmid bigger than expected after cloning?

I am currently doing restriction cloning to clone an insert (size about 4kb) into a vector with size about 3kb. I used two different REs for this cloning, and after transformation, I did colony...

12 December 2014 872 6 View

How can I solve this cloning problem?

I have two fragments and a vector, first fragment is flanked by RE A and B, second fragment is flanked by RE B and C, and the vector is flanked by A and C. Now how am I going to clone them into...

12 December 2014 1,346 13 View