PrimerPairs 1 Information (Amplicon length:2375bp)

1.Sequence information:
(1)Forward Primer:(Length28, EnzymeBamHI)
5'CGGGATCCACATTTCTTCACTTCCACAC 3'
(2)Reverse Primer:(Length28, EnzymeXhoI)
5'CCGCTCGAGTAGACTACAGTTGTTATTC 3'
2.Primer Pair Parameters:
(1)Tm(Target Annealing Region of Forward Primer):56C
Tm(Target Annealing Region of Reverse Primer):56C
Tm difference0C.
(2)GC%(Target Annealing Region of Forward Primer):40%
GC%(Target Annealing Region of Reverse Primer):33%.
(3)Forward Primer length28 nt (target annealing region length20 nt);Reverse Primer
Length28 nt(target annealing region length21 nt.)
3. Clone Strategy Information:
1) Directional cloning with double enzymes insertion!
2For the two restriction enzymes are adjacent to each other in MCS, it is recommen
ded that the plasmid pGEX-5X-1 should be dephosphorized by alkaline phosphatase
to reduce the plasmid self-ligation after double digestion. But this is not nece
ssary!
4. Restriction Enzyme Information:
(1)Promega Inc.'s BamHI(G/GATCC) and XhoI(C/TCGAG) have universal buffer:B.
(2)BamHI:
Buffer Supplied:E.
Temperature37C
Other information1.Has Star activity, No Methylation Interference.
WebSitehttp://www.promega.com/catalog/catalogproducts.aspx?categoryname=produc
tleaf_502
(3)XhoI:
Buffer Supplied:D.
Temperature37C
Other information1.No Star activityCpG methylation Interference
WebSitehttp://www.promega.com/catalog/catalogproducts.aspx?categoryname=product
leaf_597
(4) Digest Order:
[1]Gene can be double digest simultaneously in universal buffer(B) at 37C
[2]Plasmid pGEX-5X-1 can be double digest simultaneously in universal buffer
(B) at 37C