Pgem 3z
pGEM-3Z is a small, high-copy-number plasmid vector that can be used for cloning, sequencing, and in vitro transcription. It contains multiple cloning sites, a lacZ gene for blue/white screening, and T7 and SP6 RNA polymerase promoters flanking the multiple cloning site.
Lab products found in correlation
7 protocols using pgem 3z
DDR1 Mutant Generation by PCR
Quantification of LacZ Reporter Activity
In Situ Hybridization for Cytokine RNA Detection
In vitro gRNA Synthesis and CRIS-PITCh Vector Construction
Cloning eNpHR3.0 DNA into Oocyte Vectors
Chemiluminescent Quantification of lacZ Reporter
Bacterial 16S rRNA Gene Amplification
The 16S rRNA gene was amplified by PCR using universal primers 27F (5′-AGAGTTT GATCMTGGCTCAG-3′) and 1492R (5′-GGTTACCTTGTTACGACTT-3′).13 PCRs were performed in a final volume of 50 µL, containing 2 µL of DNA (50 ng/µL), 2 µL of 1 × buffer, 3 mM MgCl2, 0.2 mM each dNTP, 1 U Taq Polymerase (Fermentas, Waltham, MA, USA), 0.4 μM direct and reverse primers, and deionized water. PCR amplification was performed using a thermocycler (Mastercycler Nexus-Eppendorf) as follows: denaturation at 95° C for five minutes, followed by 30 cycles of denaturation at 94° C for 45 seconds, alignment at 56° C for 45 seconds, and extension at 72° C for 90 seconds, and a final extension of 72° C for 10 minutes. Products were separated on 1.5% agarose gels in 1 × TBE buffer stained with ethidium bromide (0.5 mg/mL) and visualized under UV light. PCR products were purified using the QIAquick PCR Purification Kit (Qiagen), according to the manufacturer's instructions. The purified PCR products were sequenced (ABI PRISM 3500 Automatic Sequencer) using the vector pGEM®-3Z (Promega, Madison, WI, USA) as a positive control.
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