What are the sequences of the primers?
SL1 is: 5′-CAG TCC AGT TAC GCT GGA GTC SR2 is: 5′-GGT CAG GTA TGA TTT AA A TGG TCA GT The pEZSeq primers are identical to the M13 Forward and Reverse Primers: Z-Rev (M13 Reverse, -48): 5′-AGC GGA TAA CAA TTT CAC ACA GGA Z-For (M13 Forward, -41): 5′-CGC CAG GGT TTT CCC AGT CAC GAC These are also listed in the Manual for each kit. 8. What are the sequences of the vectors? The vector sequences can be obtained from GenBank. The Accession numbers are as follows: pSMART-HCKan: AF532107 pSMART-LCKan: AF532106 pSMART-HCAmp: AF399742 pSMART-LCAmp: AY090111 pEZSeq-Kan: AF532108 pEZSeq-Amp: AF532109 9. How do I screen pSMART clones for inserts? In a typical ligation/transformation, there is no need to screen for inserts, since over 99% of the clones will have inserts. However, it is possible to use PCR with the SL1 and SR2 primers to confirm the presence or size of the inserts. PCR can be performed on the transformed colonies or on DNA prepared from the colonies. In addition, the cloning site of
