Avin Ee-Hwan Koh

3 Questions 5 Answers 0 Followers

Questions related from Avin Ee-Hwan Koh

Is it bad if the strongest folding Tm (secondary structure) of my primers are closely similar to that of my primers' Tm?

Would the PCR reaction be hindered if my primers can form secondary structures at a temperature that is close to the primers' melting temperature? Since the temperature difference is small, the...

30 October 2014 6,454 11 View

Should I calculate the Tm for my primers before adding in the 5' restriction sites or after?

Am I supposed to first design my primers to get the closest matching Tm and then add in the restriction sites afterwards, or are the restriction sites supposed to be added to my primers first...

20 October 2014 443 11 View

If I were to clone a gene for expression studies, should I clone the full length gene or just the CDS (with/without the start and stop codons)?

The prokaryotic gene I'm looking at has conserved DNA sequences/ sequence motifs, but do not inlcude the start and stop codons within them although the motifs are part of the CDS. So if I were to...

10 October 2014 6,885 12 View