sona_p

Great information. Do you mind if I share it on my FB page?

That's true.

Our body rejects organ grafts from other organisms.

This link might answer your question

http://www.veterinarypartner.com/Content.plx?A=1594

Kind of discrimination and generalization of your way to look at everyone from that country based on limited experience.

A study in mouse shows that female mouse prefers male with better smell than one with bad smell.

Deodorants don't exactly replicate the effect of pheromones but they increase the likely-hood of interaction with person for physical involvement.

As humans have other 100s of criteria, so just deodorants will not be able to enough to attract your mate.

Try with more DNA

If the CG are surrounded by more GC - higher TM

if GC are surrounded or interrupted by A or T - Lower TM

Another factor associated with TM

Check the total gm in the bottle. gm/liter and find out how much you need to use for 100ml

so that you can reverse calculate the required amount needed for each plate.

Perform plasmid DNA isolation that will give you all plasmids together. Then use very less amount (ngs) to transform E. coli. Grow them on selective media if they have any markers. there is a 80% chance you will be able to separately grow them and isolate them from E. coli

1 U of enzyme is the amount of enzyme that will digest 1ug of DNA in 1hr at required/optimum temperature.

I have a question before I give you any suggestion.

1) Are these two sites in your vector present in MCS of the plasmid?

If yes, then don't gel purify your vector, it will avoid loss of plasmid during the process.

2) If you are just deleting few bases from the PCR product, I would recommend skipping the gel purification step.

3) Ligate it for more than 12 hrs/over night at 4 degrees

4) Have a negative control where there is no insert added. In that way, you would get some idea about the background ligation.

Reply if you have any more questions