question on pGEM-T-easy vector system - (Jul/30/2007 )
hi! i am looking for info abt pGEM-T-easy vector bcoz im using tis vector for plasmid insertion.... n i found tis on www.promega.com....but i dont understand why it says tat it has 3`-terminal thymidine nucleotides added, is it tat tis nucleotide is important for terminal end? for stability? can anyone help me solve my question?
"The pGEM®-T and pGEM®-T Easy Vector Systems facilitate the direct cloning of PCR products into a convenient plasmid vector. The systems include pGEM®-T Vector Control DNA, T4 DNA Ligase, 2X Rapid Ligation Buffer and pGEM®-T or pGEM®-T Easy Vector. These vectors have been linearized and have had 3′-terminal thymidine nucleotides added."
errrrrrr first i would appreciate not too much abreviations... i'm older now and feel "has been"
ok pGEM "T" easy vector got poly(dT) as 3' ends.
Basically , it means that your insert should be poly(dA) tailed (10' with 1µl regular taq at 72 is perfect). The polyA tail will form appariements with the polyT of the vector, facilitating the subsequent ligation.
aha! its Fred! been reading your answers and questions before i registered to BioForum... talking about the poly(dA) tail,now i get the point...pardon me, my biology is not good...thanks
no problem, newbees are welcome and we'll help and discuss
Btw, welcome to bioforum