genetic engineering questions - not sure i understand..

[radioinahole]

I have a list of questions I have while studying for my coursecalled Introduction to genetic engineering ad I have a couple of questions ihope I can get answered. Thank you

 

 

1. When making mRNA to DNA, we use reverse transcriptase. Whenthe cDNA is made and we remove the mRNA by adding alkali. Is the complementaryDNA strand now made using DNA Polymerase I or III?

 

 

2. In genetic engineering, when a recombinant dna enters a hostcell, it either integrates, disintegrates or remains as an episome. Does thereason it disintegrates is because the host cell can detect it doesn't have thesame methylation sequence and cuts it via host controlled restriction??

 

 

3. In genetic engineering, when we want to clone a gene, I havetaken in class that we don't use PCR all the time because to use it, you haveto know the sequence of the annealing sites. I don't understand this becausewhen you want to cut a plasmid to use as a vector, should we know the sequenceanyway, to make sure we dont cut a gene or insert the foreign dna between agene hat is important for the plasmid to actually work? For example, M13 phagehas 10 genes and we shouldnt cut any of those genes otherwise we cant use it asa vector, right? So dont we know the sequence, so why cant we always use PCR?

 

 

I have a quiz coming up in 5 days and I was wondering if thereare any good sites for solving questions about ligation, restrictionendonuclease, blunt and sticky ends and pUC8 plasmids and whatnot. If you knowany sites that have answers as well as questions please let me know otherwise Iwont be able to know if I did it right or wrong. It is ok if the questions aremultiple choice. I am looking for mostly questions where they give you stickyends or the RE enzyme ad tell you if you can ligate it or how would you use itblah blah

 

Thanks for your time

 

Edited October 21, 2011 by radioinahole

[Guest Denishverma]

HI Radioinahole !

 

Your question is not much appropriate !

Not much clear -

If you can give clear display about your issue then - I am sure to help you.

 

Please repost - and give clear Image.

 

thank you !