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Rapid dephosphorylation ligation kit

Manufactured by Roche
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The Rapid dephosphorylation/ligation kit is a laboratory product designed to facilitate efficient dephosphorylation and ligation of DNA fragments. It provides the necessary reagents and protocols to perform these essential molecular biology techniques.

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Lab products found in correlation

E coli dh5α, by New England Biolabs (2 mentions) Blastidicin, by InvivoGen (2 mentions) Human puno card11 plasmid, by InvivoGen (2 mentions) Pwo dna polymerase, by Roche (2 mentions) Genelute hp plasmid maxi prep kit, by Merck Group (2 mentions)

Spelling variants of this product

Rapid Ligation Kit (12 citations) Rapid DNA Dephosphorylation and Ligation Kit (2 citations)

2 protocols using rapid dephosphorylation ligation kit

1

Generation and Validation of Modified CARD11 Constructs

Check if the same lab product or an alternative is used in the 5 most similar protocols
The human pUNO-CARD11 plasmid (Invivogen) was modified to include a 3’ 3X FLAG tag using annealed, overlapping oligonucleotides encoding the tag and inserted via BamHI (5’) and NheI (3’) overhangs. Single point mutations were introduced into the WT CARD11 construct by site-directed mutagenesis, using specific primers for linear amplification using 2x Pwo DNA polymerase (Roche) and subsequent digestion with DpnI to destroy methylated template DNA (ThermoFisher Scientific). A 42 bp fragment encoding aa183–196 duplication was generated by overlap extension PCR and ligated into a BsrG1 site in pUNO-CARD11 using a rapid dephosphorylation/ligation kit (Roche). Mutations were confirmed by Sanger sequencing. All plasmids were purified using a GenElute HP Plasmid Maxi-Prep Kit (Sigma) from transformed competent DH5α E. coli (New England Biolabs) selected with blastidicin (InvivoGen).
Ma C.A., Stinson J.R., Zhang Y., Abbott J.K., Weinreich M.A., Hauk P.J., Reynolds P.R., Lyons J.J., Nelson C.G., Ruffo E., Dorjbal B., Glauzy S., Yamakawa N., Arjunaraja S., Voss K., Stoddard J., Niemela J., Zhang Y., Rosenzweig S.D., McElwee J.J., DiMaggio T., Matthews H.F., Jones N., Stone K.D., Palma A., Oleastro M., Prieto E., Bernasconi A.R., Dubra G., Danielian S., Zaiat J., Marti M.A., Kim B., Cooper M.A., Romberg N.D., Meffre E., Gelfand E.W., Snow A.L, & Milner J.D. (2017). Germline hypomorphic CARD11 mutations in severe atopic disease. Nature genetics, 49(8), 1192-1201.
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2

Generation and Validation of Modified CARD11 Constructs

Check if the same lab product or an alternative is used in the 5 most similar protocols
The human pUNO-CARD11 plasmid (Invivogen) was modified to include a 3’ 3X FLAG tag using annealed, overlapping oligonucleotides encoding the tag and inserted via BamHI (5’) and NheI (3’) overhangs. Single point mutations were introduced into the WT CARD11 construct by site-directed mutagenesis, using specific primers for linear amplification using 2x Pwo DNA polymerase (Roche) and subsequent digestion with DpnI to destroy methylated template DNA (ThermoFisher Scientific). A 42 bp fragment encoding aa183–196 duplication was generated by overlap extension PCR and ligated into a BsrG1 site in pUNO-CARD11 using a rapid dephosphorylation/ligation kit (Roche). Mutations were confirmed by Sanger sequencing. All plasmids were purified using a GenElute HP Plasmid Maxi-Prep Kit (Sigma) from transformed competent DH5α E. coli (New England Biolabs) selected with blastidicin (InvivoGen).
Ma C.A., Stinson J.R., Zhang Y., Abbott J.K., Weinreich M.A., Hauk P.J., Reynolds P.R., Lyons J.J., Nelson C.G., Ruffo E., Dorjbal B., Glauzy S., Yamakawa N., Arjunaraja S., Voss K., Stoddard J., Niemela J., Zhang Y., Rosenzweig S.D., McElwee J.J., DiMaggio T., Matthews H.F., Jones N., Stone K.D., Palma A., Oleastro M., Prieto E., Bernasconi A.R., Dubra G., Danielian S., Zaiat J., Marti M.A., Kim B., Cooper M.A., Romberg N.D., Meffre E., Gelfand E.W., Snow A.L, & Milner J.D. (2017). Germline hypomorphic CARD11 mutations in severe atopic disease. Nature genetics, 49(8), 1192-1201.
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