How does plasmid size affect transformation efficiency?
The transformation efficiency (transformants per microgram plasmid DNA) decreased with increases of size of the DNA. The size of plasmid DNA in the range of 3.7 to 12.6 kbp did not affect the molecular efficiency (transformants per molecule input DNA).
What is the size of pBlueScript?
Plasmid: pBluescript SK (+)
| Source/Vendor: | Stratagene |
|---|---|
| Analyze: | Sequence |
| Plasmid Type: | Bacterial Expression |
| Cloning Method: | Restriction Enzyme |
| Size: | 2958 |
Why is pBlueScript a good vector?
Agilent pBlueScript II Vectors are powerful cloning vectors for a range of research applications. Featuring an extensive polylinker with 21 unique restriction enzyme recognition sites, the vectors are suitable for a range of DNA sequencing and cloning processes.
What is the pBlueScript plasmid?
The pBluescript II phagemids (plasmids with a phage origin) are cloning vectors designed to simplify commonly used cloning and sequencing procedures, including the construction of nested deletions for DNA sequencing, generation of RNA transcripts in vitro and site-specific mutagenesis and gene mapping.
What is a large plasmid size?
Large-sized plasmid (20 kb or more) isolation from.
Does insert size affect transformation efficiency?
Transformation efficiency depends on vector-insert size and bacterial strain you use. Chemical transformation (using Ca+2 for example) is worse than electroporation when you want high size inserts. Moreover, it is important to highlight that fresh competent cells work best that stored one.
What is pUC19 plasmid?
pUC19 is a small, high-copy number E. coli plasmid cloning vector, of which multiple cloning sites as shown below. The molecule is a small double-stranded circle, 2686 base pairs in length. Selection of the plasmid in E. coli is conferred by the ampicillin resistance gene.
Why is pBluescript useful?
The versatility of pBluescript vectors alleviates the need for successive subcloning. With 21 restriction sites, in two orientations, most cloned genes can be inserted directionally into the pBluescript polylinkers. If a new gene of unknown sequence is to be cloned, libraries will need to be generated for screening.
What is pUC18 plasmid?
$ 40.00. pUC18 is a commonly used plasmid cloning vector in E. coli. The molecule is a double-stranded circular DNA (2686 base pairs in length). Due to a small size pUC18 enables successful cloning of large DNA fragments.
What is the total size of the plasmid?
When the plasmid is digested with either HindIII and BamHI alone (lanes 4-5), there is a single band of 7.3 kb representing the full size of the plasmid.
How small can plasmids be?
Naturally occurring plasmids vary greatly in their physical properties. Their size can range from very small mini-plasmids of less than 1-kilobase pairs (Kbp) to very large megaplasmids of several megabase pairs (Mbp).
What is considered a large plasmid?
What can a pBluescript 2 phagemid be used for?
INTRODUCTION The pBluescript II phagemids (plasmids with a phage origin) are cloning vectors designed to simplify commonly used cloning and sequencing procedures, including the construction of nested deletions for DNA sequencing, generation of RNA transcripts in vitro and site-specific mutagenesis and gene mapping.
How is the function of pBluescript II genes validated?
To validate the function of candidate biofilm formation genes, the genes were cloned into plasmid pBluescript II SK (+) and transformed into E. coil DH5α. Biofilm production from the transformants was then assessed by phenotypic biofilm formation using Crystal Violet staining and microscopy.
How is The pGREEN plasmid system used in cloning?
This plasmid system allows any arrangement of selectable marker and reporter gene at the right and left T-DNA borders without compromising the choice of restriction sites for cloning, since the pGreen cloning sites are based on the well-known pBluescript general vector plasmids.
Can a low copy plasmid be used as a vector?
A low copy plasmid or even a bacterial artificial chromosome (BAC) vector could perhaps solve this problem (She, 2003;Shizuya et al., 1992), for instance, the respiratory syncytial virus was successfully rescued from a BAC vector (Hotard et al., 2012).