Mutagenesis:Article Title: m 3 C32 tRNA modification controls serine codon-biased mRNA translation, cell cycle, and DNA-damage response.
Article Snippet: Next, the digested insert was ligated into the cut pCMV2Flag-empty vector using T4 DNA ligase (NEB, #M0202). .. All pCMV2Flag-METTL2A and pCMV2-Flag-METTL6 mutation constructs were generated by using a Q5 site-directed mutagenesis kit (NEB, #E0554) except pCMV2-Flag-METTL2A-dC. pCMV2-Flag-METTL2A-dC was generated by subcloning the cDNA of METTL2A with a deletion of amino acids 287-378 into the pCMV2-Flag-empty vector. .. For the construction of METTL2A/2B and/or METTL6 sgRNA vectors for CRISPR-Cas9 knockout, sgRNAs were designed using the CRISPR design website (http://crispr.mit.edu/). sgRNA specificity was checked by using the Cas-OFFinder website (http://www.rgenome.netcas-offinder/).
Article Title: m 3 C32 tRNA modification controls serine codon-biased mRNA translation, cell cycle, and DNA-damage response
Article Snippet: Next, the digested insert was ligated into the cut pCMV2-Flag-empty vector using T4 DNA ligase (NEB, #M0202). .. All pCMV2-Flag-METTL2A and pCMV2-Flag-METTL6 mutation constructs were generated by using a Q5 site-directed mutagenesis kit (NEB, #E0554) except pCMV2-Flag-METTL2A-dC. pCMV2-Flag-METTL2A-dC was generated by subcloning the cDNA of METTL2A with a deletion of amino acids 287-378 into the pCMV2-Flag-empty vector. .. For the construction of METTL2A/2B and/or METTL6 sgRNA vectors for CRISPR-Cas9 knockout, sgRNAs were designed using the CRISPR design website ( http://crispr.mit.edu/ ). sgRNA specificity was checked by using the Cas-OFFinder website ( http://www.rgenome.netcas-offinder/ ).
Construct:Article Title: m 3 C32 tRNA modification controls serine codon-biased mRNA translation, cell cycle, and DNA-damage response.
Article Snippet: Next, the digested insert was ligated into the cut pCMV2Flag-empty vector using T4 DNA ligase (NEB, #M0202). .. All pCMV2Flag-METTL2A and pCMV2-Flag-METTL6 mutation constructs were generated by using a Q5 site-directed mutagenesis kit (NEB, #E0554) except pCMV2-Flag-METTL2A-dC. pCMV2-Flag-METTL2A-dC was generated by subcloning the cDNA of METTL2A with a deletion of amino acids 287-378 into the pCMV2-Flag-empty vector. .. For the construction of METTL2A/2B and/or METTL6 sgRNA vectors for CRISPR-Cas9 knockout, sgRNAs were designed using the CRISPR design website (http://crispr.mit.edu/). sgRNA specificity was checked by using the Cas-OFFinder website (http://www.rgenome.netcas-offinder/).
Article Title: m 3 C32 tRNA modification controls serine codon-biased mRNA translation, cell cycle, and DNA-damage response
Article Snippet: Next, the digested insert was ligated into the cut pCMV2-Flag-empty vector using T4 DNA ligase (NEB, #M0202). .. All pCMV2-Flag-METTL2A and pCMV2-Flag-METTL6 mutation constructs were generated by using a Q5 site-directed mutagenesis kit (NEB, #E0554) except pCMV2-Flag-METTL2A-dC. pCMV2-Flag-METTL2A-dC was generated by subcloning the cDNA of METTL2A with a deletion of amino acids 287-378 into the pCMV2-Flag-empty vector. .. For the construction of METTL2A/2B and/or METTL6 sgRNA vectors for CRISPR-Cas9 knockout, sgRNAs were designed using the CRISPR design website ( http://crispr.mit.edu/ ). sgRNA specificity was checked by using the Cas-OFFinder website ( http://www.rgenome.netcas-offinder/ ).
Generated:Article Title: m 3 C32 tRNA modification controls serine codon-biased mRNA translation, cell cycle, and DNA-damage response.
Article Snippet: Next, the digested insert was ligated into the cut pCMV2Flag-empty vector using T4 DNA ligase (NEB, #M0202). .. All pCMV2Flag-METTL2A and pCMV2-Flag-METTL6 mutation constructs were generated by using a Q5 site-directed mutagenesis kit (NEB, #E0554) except pCMV2-Flag-METTL2A-dC. pCMV2-Flag-METTL2A-dC was generated by subcloning the cDNA of METTL2A with a deletion of amino acids 287-378 into the pCMV2-Flag-empty vector. .. For the construction of METTL2A/2B and/or METTL6 sgRNA vectors for CRISPR-Cas9 knockout, sgRNAs were designed using the CRISPR design website (http://crispr.mit.edu/). sgRNA specificity was checked by using the Cas-OFFinder website (http://www.rgenome.netcas-offinder/).
Article Title: m 3 C32 tRNA modification controls serine codon-biased mRNA translation, cell cycle, and DNA-damage response
Article Snippet: Next, the digested insert was ligated into the cut pCMV2-Flag-empty vector using T4 DNA ligase (NEB, #M0202). .. All pCMV2-Flag-METTL2A and pCMV2-Flag-METTL6 mutation constructs were generated by using a Q5 site-directed mutagenesis kit (NEB, #E0554) except pCMV2-Flag-METTL2A-dC. pCMV2-Flag-METTL2A-dC was generated by subcloning the cDNA of METTL2A with a deletion of amino acids 287-378 into the pCMV2-Flag-empty vector. .. For the construction of METTL2A/2B and/or METTL6 sgRNA vectors for CRISPR-Cas9 knockout, sgRNAs were designed using the CRISPR design website ( http://crispr.mit.edu/ ). sgRNA specificity was checked by using the Cas-OFFinder website ( http://www.rgenome.netcas-offinder/ ).
Subcloning:Article Title: m 3 C32 tRNA modification controls serine codon-biased mRNA translation, cell cycle, and DNA-damage response.
Article Snippet: Next, the digested insert was ligated into the cut pCMV2Flag-empty vector using T4 DNA ligase (NEB, #M0202). .. All pCMV2Flag-METTL2A and pCMV2-Flag-METTL6 mutation constructs were generated by using a Q5 site-directed mutagenesis kit (NEB, #E0554) except pCMV2-Flag-METTL2A-dC. pCMV2-Flag-METTL2A-dC was generated by subcloning the cDNA of METTL2A with a deletion of amino acids 287-378 into the pCMV2-Flag-empty vector. .. For the construction of METTL2A/2B and/or METTL6 sgRNA vectors for CRISPR-Cas9 knockout, sgRNAs were designed using the CRISPR design website (http://crispr.mit.edu/). sgRNA specificity was checked by using the Cas-OFFinder website (http://www.rgenome.netcas-offinder/).
Article Title: m 3 C32 tRNA modification controls serine codon-biased mRNA translation, cell cycle, and DNA-damage response
Article Snippet: Next, the digested insert was ligated into the cut pCMV2-Flag-empty vector using T4 DNA ligase (NEB, #M0202). .. All pCMV2-Flag-METTL2A and pCMV2-Flag-METTL6 mutation constructs were generated by using a Q5 site-directed mutagenesis kit (NEB, #E0554) except pCMV2-Flag-METTL2A-dC. pCMV2-Flag-METTL2A-dC was generated by subcloning the cDNA of METTL2A with a deletion of amino acids 287-378 into the pCMV2-Flag-empty vector. .. For the construction of METTL2A/2B and/or METTL6 sgRNA vectors for CRISPR-Cas9 knockout, sgRNAs were designed using the CRISPR design website ( http://crispr.mit.edu/ ). sgRNA specificity was checked by using the Cas-OFFinder website ( http://www.rgenome.netcas-offinder/ ).
Plasmid Preparation:Article Title: m 3 C32 tRNA modification controls serine codon-biased mRNA translation, cell cycle, and DNA-damage response.
Article Snippet: Next, the digested insert was ligated into the cut pCMV2Flag-empty vector using T4 DNA ligase (NEB, #M0202). .. All pCMV2Flag-METTL2A and pCMV2-Flag-METTL6 mutation constructs were generated by using a Q5 site-directed mutagenesis kit (NEB, #E0554) except pCMV2-Flag-METTL2A-dC. pCMV2-Flag-METTL2A-dC was generated by subcloning the cDNA of METTL2A with a deletion of amino acids 287-378 into the pCMV2-Flag-empty vector. .. For the construction of METTL2A/2B and/or METTL6 sgRNA vectors for CRISPR-Cas9 knockout, sgRNAs were designed using the CRISPR design website (http://crispr.mit.edu/). sgRNA specificity was checked by using the Cas-OFFinder website (http://www.rgenome.netcas-offinder/).
Article Title: m 3 C32 tRNA modification controls serine codon-biased mRNA translation, cell cycle, and DNA-damage response
Article Snippet: Next, the digested insert was ligated into the cut pCMV2-Flag-empty vector using T4 DNA ligase (NEB, #M0202). .. All pCMV2-Flag-METTL2A and pCMV2-Flag-METTL6 mutation constructs were generated by using a Q5 site-directed mutagenesis kit (NEB, #E0554) except pCMV2-Flag-METTL2A-dC. pCMV2-Flag-METTL2A-dC was generated by subcloning the cDNA of METTL2A with a deletion of amino acids 287-378 into the pCMV2-Flag-empty vector. .. For the construction of METTL2A/2B and/or METTL6 sgRNA vectors for CRISPR-Cas9 knockout, sgRNAs were designed using the CRISPR design website ( http://crispr.mit.edu/ ). sgRNA specificity was checked by using the Cas-OFFinder website ( http://www.rgenome.netcas-offinder/ ).
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