The pDrive Cloning Vector (see figure " pDrive Cloning Vector") provides highly efficient cloning of PCR products through UA hybridization. The vector is supplied in a linear form with a U overhang at each 3' end, which hybridizes with high specificity to the A overhang of PCR products generated by Taq and other nonproofreading DNA polymerases. The pDrive Cloning Vector (see figure "pDrive Cloning Vector") has a number of useful features designed to facilitate analysis of cloned PCR products. These include a large number of unique restriction enzyme recognition sites, universal sequencing primer sites, and promoters for in vitro transcription. In addition, the vector allows both ampicillin and kanamycin selection, as well as blue/white screening of recombinant colonies.
PCR products are efficiently cloned into the pDrive Cloning Vector in less time than is required for TA-based cloning vectors (see figure "Highly specific cloning with a shorter ligation time of 30 min"). Furthermore, as T is the most likely base to hybridize to noncomplementary bases (i.e., G, C, and T), vectors with a T overhang are more likely to self-anneal or to clone primers or annealed primers, leading to an increased number of false-positive colonies. In contrast, the higher cloning efficiency of the pDrive Cloning Vector indicates that U has a lower tolerance for nonspecific base pairing.
The Ligation Master Mix, supplied in a convenient premixed format, is specifically designed to provide optimal hybridization conditions for efficient cloning.
The use of QIAGEN EZ Competent Cells in the QIAGEN PCR Cloningplus Kit makes the cloning procedure even faster and more convenient. Transformed cells are usually incubated in SOC medium to recover and to have time to express antibiotic resistance. In contrast, QIAGEN EZ Competent Cells do not require this time-consuming recovery incubation to achieve high-efficiency transformation (>108 colony forming units (CFU) per microgram DNA; see figure "Transformation without recovery incubation").
Components included with QIAGEN PCR Cloning Kits
|Component ||QIAGEN PCR Cloning Kit ||QIAGEN PCR Cloningplus Kit ||Concentration |
|pDrive Cloning Vector ||+ ||+ ||50 ng/µl |
|Ligation Master Mix ||+ ||+ ||2x solution |
|Distilled water ||+ ||+ ||– |
|QIAGEN EZ Competent Cells ||– ||+ ||– |
|SOC medium ||– ||+ ||– |