Make your target specific gRNA lentivectors with desired markers!
pLenti-gRNA-U6 / H1- Cloning Kit is designed for cloning of double-strand DNA encoding your desired gRNA sequence for CRISPR genomic gene editing. Once transcribed, the target specific gRNA will guild CAS9 enzyme to cut the genomic DNA at desired loci.
To make your target specific gRNA lentivector, two synthetic gRNA oligonucleotides are first annealed to form the double stand DNA duplex which is then cloned into the provided, linear pLenti-gRNA-U6 / H1 lentivector via T4 enzyme ligation.
1) Flexibility to select desired promoter: H1 or U6;
2) Flexibility to choose desired selection marker: antibiotic or antibiotic-fluorescent fusion dual marker;
3) Easy and high efficiency cloning reaction: with provided pre-cut, ready to use, linear gRNA lentivector, you do NOT need the tedious, yet often troublesome lentivector backbone preparation. The Cloning positive rate close to 100%.
4) Everything is included in this kit, enough for generating 10 gRNA lentivectors;
5) Optionally used as tetracycline inducible gRNA transcription (for H1 promoter only);
This product is the lentivector version having the gRNA driven by a constitutive an optional inducible human H1 promoter with a RFP-Puromycin dual selection marker under Rsv promoter. Please see Product Manual for details.
pLenti-H1-(RFP-Puro) linear vector: 10 ul (10rxn, 1ul/rxn)
10x gRNA oligo annealing solution: 25 ul
10X ligation buffer: 20 ul
T4 ligase enzyme: 10 ul (10rxn, 1ul/rxn)
Cloning control insert: annealed Neg-gRNA duplex: 10 ul (10rxn, 1ul/rxn)
Sequencing primer: 10 ul (10xn, 1ul/rxn)
DH5a chemical competent cells: 10 vials (10xn, 1 vial/rxn)
The generated gRNA lentivector can be used for transfection or electroporation based CRISPR editing, or used for gRNA lentivirus production and then use the gRNA lentivirus for CRISPR editing.