RPA 8 Cloning and CRISPR
If you want to clone Gene 1 into the puC19 plasmid so it is in the same orientation as the promoter, which of the following enzymes could you use to do this? Note your amp gene must be compete Note: ScaI, and HincII are blunt end digesters. Hind III is a sticky end digester.
HindIII and HincII to cut the plasmid and HindIII and ScaI to cut gene 1
If you want to clone Gene 1 into the puC19 plasmid so it is in the same orientation as the promoter, which of the following enzymes could you use to do this? Note: your amp gene must be complete. Note: ScaI, and SmaI are blunt end digesters. KpnI is a sticky end digester.
Kpn and SmaI to cut the plasmid and KpnI and ScaI to cut gene 1
You have the following restriction enzymes with the indicated recognition sequence. The ^ indicates the site of cleavage. If you digest a plasmid with Pov II, can you ligate it to a gene of interest that has been Restriction enzyme digested with Bam HI? BamHI G^GATCC CCTAG^G PovII CAG^CTG GTC^GAC
No
You have the following restriction enzymes with the indicated recognition sequence. The ^ indicates the site of cleavage. If you digest a plasmid with Pov II, can you ligate it to a gene of interest that has been restriction enzyme digested with Eco RI? Eco RI G^AATTC CTTAA^G PovII CAG^CTG GTC^GAC
No
You have the following restriction enzymes with the indicated recognition sequence. The ^ indicates the site of cleavage. If you digest a plasmid with Pov II, can you ligate it to a gene of interest that has been restriction enzyme digested with Psp AI? PspAI A^CCGGT TGGCC^A PovII CAG^CTG GTC^GAC
No
You have the following restriction enzymes with the indicated recognition sequence. The ^ indicates the site of cleavage. If you digest a plasmid with Sma I, can you ligate it to a gene of interest that has been restriction enzyme digested with Psp AI? Sma I CCC^GGG GGG^CCC PspAI A^CCGGT TGGCC^A
No
You have the following restriction enzymes with the indicated recognition sequence. The ^ indicates the site of cleavage. If you digest a plasmid with SmaI, can you ligate it to a gene of interest that has been Restriction enzyme digested with Bam HI? Sma I CCC^GGG GGG^CCC BamHI G^GATCC CCTAG^G
No
You have the following restriction enzymes with the indicated recognition sequence. The ^ indicates the site of cleavage. If you digest a plasmid with SmaI, can you ligate it to a gene of interest that has been restriction enzyme digested with Xma I? Xma I C^CCGGG GGGCC^C Sma I CCC^GGG GGG^CCC
No
You have the following restriction enzymes with the indicated recognition sequence. The ^ indicates the site of cleavage. You are setting up a cloning assay you digest your cloning vector with BAMHI and you digest your gene of interest with Eco RI. Can you ligate together these 2 pieces of DNA? BamHI G^GATCC CCTAG^G Eco RI G^AATTC CTTAA^G
No
You have the following restriction enzymes with the indicated recognition sequence. The ^ indicates the site of cleavage. You are setting up a cloning assay you digest your cloning vector with Psp AI and you digest your gene of interest with BAMHI. Can you ligate together these 2 pieces of DNA? BamHI G^GATCC CCTAG^G PspAI A^CCGGT TGGCC^A
No
You have the following restriction enzymes with the indicated recognition sequence. The ^ indicates the site of cleavage. You are setting up a cloning assay you digest your cloning vector with Psp AI and you digest your gene of interest with Eco RI. Can you ligate together these 2 pieces of DNA? PspAI A^CCGGT TGGCC^A Eco RI G^AATTC CTTAA^G
No
You have the following restriction enzymes with the indicated recognition sequence. The ^ indicates the site of cleavage. You are setting up a cloning assay you digest your cloning vector with XmaI and you digest your gene of interest with Bam HI. Can you ligate together these 2 pieces of DNA? Xma I C^CCGGG GGGCC^C BamHI G^GATCC CCTAG^G
No
You have the following restriction enzymes with the indicated recognition sequence. The ^ indicates the site of cleavage. You are setting up a cloning assay you digest your cloning vector with XmaI and you digest your gene of interest with Eco RI. Can you ligate together these 2 pieces of DNA? Xma I C^CCGGG GGGCC^C Eco RI G^AATTC CTTAA^G
No
If you want to clone Gene 1 into the puC19 plasmid so it is in the same orientation as the promoter, which of the following enzymes could you use to do this? Note your amp gene must be compete Note: ScaI, and HincII are blunt end digesters. PstI is a sticky end digester.
PstI and HincII to cut the plasmid and PstI and ScaI to cut gene 1
If you want to clone Gene 1 into the puC19 plasmid so it is in the same orientation as the promoter, which of the following enzymes could you use to do this? Note: ScaI, and SmaI are blunt end digesters. SacI is a sticky end digester.
SacI and SmaI to cut the plasmid and SacI and ScaI to cut gene 1
You have the following restriction enzymes with the indicated recognition sequence. The ^ indicates the site of cleavage. You are setting up a cloning assay you digest your cloning vector with PovII and you digest your gene of interest with Alu I. Can you ligate together these 2 pieces of DNA? PovII CAG^CTG GTC^GAC AluI AG^CT TC^GA
Yes
You have the following restriction enzymes with the indicated recognition sequence. The ^ indicates the site of cleavage. You are setting up a cloning assay you digest your cloning vector with PovII and you digest your gene of interest with Dra I. Can you ligate together these 2 pieces of DNA? PovII CAG^CTG GTC^GAC DraI TTT^AAA AAA^TTT
Yes
You have the following restriction enzymes with the indicated recognition sequence. The ^ indicates the site of cleavage. You are setting up a cloning assay you digest your cloning vector with PovII and you digest your gene of interest with SmaI. Can you ligate together these 2 pieces of DNA? PovII CAG^CTG GTC^GAC Sma I CCC^GGG GGG^CCC
Yes
You have the following restriction enzymes with the indicated recognition sequence. The ^ indicates the site of cleavage. You are setting up a cloning assay you digest your cloning vector with Sma I and you digest your gene of interest with Afe I. Can you ligate together these 2 pieces of DNA? Sma I CCC^GGG GGG^CCC AfeI AGC^GCT TCG^CGA
Yes
You have the following restriction enzymes with the indicated recognition sequence. The ^ indicates the site of cleavage. You are setting up a cloning assay you digest your cloning vector with Sma I and you digest your gene of interest with Alu I. Can you ligate together these 2 pieces of DNA? AluI AG^CT TC^GA Sma I CCC^GGG GGG^CCC
Yes
You have the following restriction enzymes with the indicated recognition sequence. The ^ indicates the site of cleavage. You are setting up a cloning assay you digest your cloning vector with SmaI and you digest your gene of interest with Dra I. Can you ligate together these 2 pieces of DNA? DraI TTT^AAA AAA^TTT Sma I CCC^GGG GGG^CCC
Yes
You have the following restriction enzymes with the indicated recognition sequence. The ^ indicates the site of cleavage. If you digest a plasmid with BamHI, can you ligate it to a gene of interest that has been Restriction enzyme digested with BcII? BamHI G^GATCC CCTAG^G BclI T^GATCA ACTAG^T
Yes
You have the following restriction enzymes with the indicated recognition sequence. The ^ indicates the site of cleavage. If you digest a plasmid with BamHI, can you ligate it to a gene of interest that has been Restriction enzyme digested with BgIII? BamHI G^GATCC CCTAG^G BglII A^GATCT TCTAG^A
Yes
You have the following restriction enzymes with the indicated recognition sequence. The ^ indicates the site of cleavage. If you digest a plasmid with BcII, can you ligate it to a gene of interest that has been Restriction enzyme digested with BgIII? BglII A^GATCT TCTAG^A BclI T^GATCA ACTAG^T
Yes
You have the following restriction enzymes with the indicated recognition sequence. The ^ indicates the site of cleavage. If you digest a plasmid with XmaI, can you ligate it to a gene of interest that has been Restriction enzyme digested with PspAI? Xma I C^CCGGG GGGCC^C PspAI A^CCGGT TGGCC^A
Yes
You have the following restriction enzymes with the indicated recognition sequence. The ^ indicates the site of cleavage. You are setting up a cloning assay you digest your cloning vector with PovII and you digest your gene of interest with Afe I. Can you ligate together these 2 pieces of DNA? PovII CAG^CTG GTC^GAC AfeI AGC^GCT TCG^CGA
Yes
You have the following restriction enzymes with the indicated recognition sequence. The ^ indicates the site of cleavage. You are setting up a cloning assay you digest your cloning vector with PovII and you digest your gene of interest with Afe I. Can you ligate together these 2 pieces of DNA? PovII CAG^CTG GTC^GAC AfeI AGC^GCT TCG^CGA
Yes
