Log In Sign up
The largest database of trusted experimental protocols

Bsiwi

Manufactured by Takara Bio
Visit Supplier
Sourced in United States, China

BsiWI is a type II restriction endonuclease that recognizes and cleaves the palindromic DNA sequence 5'-CCGC(G/C)G-3'. This enzyme is commonly used in molecular biology applications such as DNA cloning, genetic engineering, and DNA analysis.

Automatically generated - may contain errors

Lab products found in correlation

Xhoi, by Takara Bio (5 mentions) Pezx fr02 vector, by GeneCopoeia (4 mentions) Bsiwi, by New England Biolabs (2 mentions) E2 crimson template, by Takara Bio (2 mentions) Buffer 3, by New England Biolabs (2 mentions) Pcd2.1 cir, by Geneseed (1 mentions) Pezx fr02, by GeneCopoeia (1 mentions) Bamhi, by Takara Bio (1 mentions)

7 protocols using bsiwi

1

miR-148a-3p Binding Site Characterization

Check if the same lab product or an alternative is used in the 5 most similar protocols
Fragments of miR-148a-3p, including the binding site of Meox2, were amplified and inserted into pEZX-FR02 vectors (GeneCopoeia, United States) at the 3′ end of the Firefly Luciferase gene using restriction enzymes BsiWI and XhoI (TaKaRa, Dalian, China) and T4 DNA ligase (pEZX-Meox2-WT). Mutant pEZX-Meox2-MT was generated by mutating complementary to the seed region of miR-148a-3p using mutagenic primers. All constructs were verified by sequence analysis.
Yin H., He H., Cao X., Shen X., Han S., Cui C., Zhao J., Wei Y., Chen Y., Xia L., Wang Y., Li D, & Zhu Q. (2020). MiR-148a-3p Regulates Skeletal Muscle Satellite Cell Differentiation and Apoptosis via the PI3K/AKT Signaling Pathway by Targeting Meox2. Frontiers in Genetics, 11, 512.
+ Open protocol
+ Expand
2

Investigating miR-3525 Regulation of PDLIM3

Check if the same lab product or an alternative is used in the 5 most similar protocols
Fragments of gga-miR-3525, including the binding site of PDLIM3 (150 bp upstream and downstream of the binding site), were amplified and inserted into pEZX-FR02 vectors (GeneCopoeia, USA) at the 3’ end of the Firefly Luciferase gene using restriction enzymes BsiWI and XhoI (TaKaRa, Dalian, China) and T4 DNA ligase (pEZX-PDLIM3-WT). Mutant pEZX-PDLIM3-MT was generated by mutating complementary to the seed region of gga-miR-3525 using mutagenic primers. All constructs were verified by sequence analysis.
Yin H., Zhao J., He H., Chen Y., Wang Y., Li D, & Zhu Q. (2020). Gga-miR-3525 Targets PDLIM3 through the MAPK Signaling Pathway to Regulate the Proliferation and Differentiation of Skeletal Muscle Satellite Cells. International Journal of Molecular Sciences, 21(15), 5573.
+ Open protocol
+ Expand
3

Cloning miR-9-5p binding site in Luciferase

Check if the same lab product or an alternative is used in the 5 most similar protocols
Fragments of miR-9-5p, including the binding site of IGF2BP3, were amplified and inserted into pEZX-FR02 vectors (GeneCopoeia, Amaranth Drive Germantown, Maryland, USA) at the 3’ end of the Firefly Luciferase gene using restriction enzymes BsiWI and XhoI (TaKaRa) and T4 DNA ligase (pEZX- IGF2BP3-WT). Mutant pEZX- IGF2BP3-MT was generated by mutating complementary to the seed region of miR-9-5p using mutagenic primers. All constructs were verified by sequence analysis.
Yin H., He H., Shen X., Zhao J., Cao X., Han S., Cui C., Chen Y., Wei Y., Xia L., Wang Y., Li D, & Zhu Q. (2020). miR-9-5p Inhibits Skeletal Muscle Satellite Cell Proliferation and Differentiation by Targeting IGF2BP3 through the IGF2-PI3K/Akt Signaling Pathway. International Journal of Molecular Sciences, 21(5), 1655.
+ Open protocol
+ Expand
4

Generating Luciferase Reporter Constructs

Check if the same lab product or an alternative is used in the 5 most similar protocols
The linear sequence of circTMTC1 was synthesized and cloned into pCD2.1-ciR (Geneseed Biotech, Guangzhou, China) according to the manufacturer's protocol using the KpnI and BamHI (TaKaRa) restriction sites (pCD2.1-ciRcTMTC1). The fragment of the circTMTC1 including binding site of miR-128-3p, was amplified and inserted into Dual luciferase reporter vector (pEZX-FR02) (GeneCopoeia, Rockville, MD, USA) at the 3′ end of Firefly Luciferase gene using restriction enzymes BsiWI and XhoI (TaKaRa) and T4 DNA ligase (pEZX-circTMTC1-WT). The mutant pEZX-circTMTC1-MT was generated by mutating complementary to the seed region of the miR-128-3p using mutagenic primer. pEZX-MSTN-3'UTR-WT vector and pEZX-MSTN-3'UTR-MT vector were constructed using the same method. All constructs were verified by sequencing.
Small interfering RNA (siRNA) overlap junction sites of circTMTC1 and miR-128-3p mimics/inhibitor (Supplementary Table. S2) were synthesized by GenePharma Co. Ltd (Shanghai, China).
Shen X., Liu Z., Cao X., He H., Han S., Chen Y., Cui C., Zhao J., Li D., Wang Y., Zhu Q, & Yin H. (2019). Circular RNA profiling identified an abundant circular RNA circTMTC1 that inhibits chicken skeletal muscle satellite cell differentiation by sponging miR-128-3p. International Journal of Biological Sciences, 15(10), 2265-2281.
+ Open protocol
+ Expand
5

Cloning and Mutagenesis of miR-200a-3p Binding Site

Check if the same lab product or an alternative is used in the 5 most similar protocols
Fragments of miR-200a-3p, including the binding site of TGF-β2, were amplified and inserted into pEZX-FR02 vectors (GeneCopoeia, Amaranth Drive Germantown, Maryland, USA) at the 3′ end of the Firefly Luciferase gene using restriction enzymes BsiWI and XhoI (TaKaRa) and T4 DNA ligase (pEZX-TGF-β2-WT). Mutant pEZX-TGF-β2-MT was generated by mutating complementary to the seed region of miR-200a-3p using mutagenic primers. All constructs were verified by sequence analysis.
Yin H., He H., Shen X., Tang S., Zhao J., Cao X., Han S., Cui C., Chen Y., Wei Y., Wang Y., Li D, & Zhu Q. (2020). MicroRNA Profiling Reveals an Abundant miR-200a-3p Promotes Skeletal Muscle Satellite Cell Development by Targeting TGF-β2 and Regulating the TGF-β2/SMAD Signaling Pathway. International Journal of Molecular Sciences, 21(9), 3274.
+ Open protocol
+ Expand
6

Cloning of E2-Crimson Template into lentiGuide-Puro

Check if the same lab product or an alternative is used in the 5 most similar protocols
E2-Crimson template (Clontech) was PCR amplified to add BsiWI (5′) and MluI (3′) restriction sites for cloning purposes using the following conditions: KOD buffer (1×), MgSO4 (1.5 mM), dNTPs (0.2 mM each), forward primer (0.3 μM; GGCCGGCCCGTACGcgtacgGCCACCATGGATAGCACTGAGAACGTCATCAAGCCCTT), reverse primer (0.3 μM; GGCCGGCCacgcgtCTACTGGAACAGGTGGTGGCGGGCCT), and KOD Hot Start DNA Polymerase (0.02 U/μL). KOD PCR reaction used the following cycling conditions: 95°C for 2 minutes; 50 cycles of 95°C for 20 seconds, 60°C for 20 seconds, and 70°C for 30 seconds; 60°C for 5 minutes. PCR products were purified (QIAquick PCR Purification Kit) and cloned with Zero Blunt PCR cloning kit. Cloned products and lentiGuide-puro were separately digested with BsiWI (New England Biolabs) and MluI (New England Biolabs) in 1× Buffer 3.1 at 37°C (New England Biolabs). Digest of lentiGuide-Puro (Addgene plasmid ID 52963) was performed to remove the puromycin cassette. Then digested PCR product was ligated into the lentiGuide backbone.
Canver M.C., Smith E.C., Sher F., Pinello L., Sanjana N.E., Shalem O., Chen D.D., Schupp P.G., Vinjamur D.S., Garcia S.P., Luc S., Kurita R., Nakamura Y., Fujiwara Y., Maeda T., Yuan G.C., Feng Z., Orkin S.H, & Bauer D.E. (2015). BCL11A enhancer dissection by Cas9-mediated in situ saturating mutagenesis. Nature, 527(7577), 192-197.
+ Open protocol
+ Expand
7

Cloning of E2-Crimson Template into lentiGuide-Puro

Check if the same lab product or an alternative is used in the 5 most similar protocols
E2-Crimson template (Clontech) was PCR amplified to add BsiWI (5′) and MluI (3′) restriction sites for cloning purposes using the following conditions: KOD buffer (1×), MgSO4 (1.5 mM), dNTPs (0.2 mM each), forward primer (0.3 μM; GGCCGGCCCGTACGcgtacgGCCACCATGGATAGCACTGAGAACGTCATCAAGCCCTT), reverse primer (0.3 μM; GGCCGGCCacgcgtCTACTGGAACAGGTGGTGGCGGGCCT), and KOD Hot Start DNA Polymerase (0.02 U/μL). KOD PCR reaction used the following cycling conditions: 95°C for 2 minutes; 50 cycles of 95°C for 20 seconds, 60°C for 20 seconds, and 70°C for 30 seconds; 60°C for 5 minutes. PCR products were purified (QIAquick PCR Purification Kit) and cloned with Zero Blunt PCR cloning kit. Cloned products and lentiGuide-puro were separately digested with BsiWI (New England Biolabs) and MluI (New England Biolabs) in 1× Buffer 3.1 at 37°C (New England Biolabs). Digest of lentiGuide-Puro (Addgene plasmid ID 52963) was performed to remove the puromycin cassette. Then digested PCR product was ligated into the lentiGuide backbone.
Canver M.C., Smith E.C., Sher F., Pinello L., Sanjana N.E., Shalem O., Chen D.D., Schupp P.G., Vinjamur D.S., Garcia S.P., Luc S., Kurita R., Nakamura Y., Fujiwara Y., Maeda T., Yuan G.C., Feng Z., Orkin S.H, & Bauer D.E. (2015). BCL11A enhancer dissection by Cas9-mediated in situ saturating mutagenesis. Nature, 527(7577), 192-197.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!