Shrimp alkaline phosphatase

Manufactured by Cytiva

Sourced in United Kingdom

Shrimp alkaline phosphatase is an enzyme that catalyzes the removal of phosphate groups from various molecules, including nucleic acids and proteins. It is commonly used in molecular biology applications to remove unwanted phosphate groups from DNA and RNA samples, facilitating subsequent enzymatic manipulations.

Automatically generated - may contain errors

Lab products found in correlation

Exonuclease 1, by Cytiva (3 mentions) Topo ta cloning kit, by Thermo Fisher Scientific (1 mentions) M13 reverse primer, by Thermo Fisher Scientific (1 mentions) Qiaquick pcr purification kit, by Qiagen (1 mentions)

4 protocols using shrimp alkaline phosphatase

Mitochondrial COI Amplification and Sequencing

Check if the same lab product or an alternative is used in the 5 most similar protocols

The cytochrome oxidase subunit 1 (COI) region of the mitochondrial genome was amplified by polymerase chain reaction (PCR) using the primers and thermocycler conditions described by Campbell et al. (2005 (link)). PCR products were verified by electrophoresis on a 1.5% agarose gel and purified using Exonuclease I (EXO, Amersham Biosciences cat. #E70073X, 10 U/ml) and Shrimp Alkaline Phosphatase (SAP, Amersham Biosciences cat. #E70092X 1 U/ml). An EXOSAP solution was created with 78 µl ddH₂O, 2 µl EXO, and 20 µl SAP, and then, 2 µl of the mixture was added to each PCR product to remove primers, dNTPs, and other impurities, and were Sanger sequenced by Eton Bioscience (www.etonbio.com, San Diego, California) using the forward primer (Campbell et al., 2005 (link)).

Bucholz J.R., Sard N.M., VanTassel N.M., Lozier J.D., Morris T.J., Paquet A, & Zanatta D.T. (2022). RAD‐tag and mitochondrial DNA sequencing reveal the genetic structure of a widespread and regionally imperiled freshwater mussel, Obovaria olivaria (Bivalvia: Unionidae). Ecology and Evolution, 12(1), e8560.

+ Open protocol

+ Expand

Variant Identification via FSSCP and Sequencing

Check if the same lab product or an alternative is used in the 5 most similar protocols

Samples in which a variant was identified by FSSCP were re-amplified using unlabeled PCR primers. PCR products were prepared for sequencing by treatment of 5 µl of PCR product with 2 U of shrimp alkaline phosphatase and 10 U of exonuclease I (Amersham Pharmacia, Chalfont St Giles, UK) at 37°C for 30 minutes and then 80°C for 15 minutes. Sequencing reactions of 10 µl were carried out using 1–3 µl of prepared PCR product, 1.6 picomoles of primer, BigDye ready reaction mix (Applied Biosystems) version 2 diluted 1∶2 with Half Big Dye reagent (Genetix, http://www.genetix.com/). Cycle sequencing conditions were 25 cycles of 96°C for 10 seconds, 50°C for 5 seconds and 60°C for 4 minutes. Unincorporated nucleotides and primers were removed by ethanol precipitation. Sequencing products were resuspended in 10 µl of formamide and run on 310 or 3100 genetic analysers (Applied Biosystems) using POP6 and 36 cm well-to-read capillaries. Data analysis was carried out by visual inspection of electropherograms.

Zauber P., Bishop T., Taylor C., Sabbath-Solitare M., Marotta S, & Tomlinson I. (2014). Colorectal Tumors from APC*I1307K Carriers Principally Harbor Somatic APC Mutations outside the A8 Tract. PLoS ONE, 9(1), e84498.

+ Open protocol

+ Expand

Cloning and Sequencing Feline Oral Microbiome

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Purified DNA was cloned using a TOPO TA Cloning Kit (Invitrogen, Carlsbad, CA) following the manufacturer’s instructions. Transformation was done using competent E. coli TOP10 cells provided by the manufacturer. The transformed cells were plated onto Luria-Bertani agar plates supplemented with kanamycin (50 µg/ml) and incubated overnight at 37°C. Each white colony was placed into a tube containing 40 µL of 10 mM Tris-HCl pH 8.0. One µL of the cell suspension was used directly as template for PCR with an M13 (−21) forward primer and an M13 reverse primer (Invitrogen). The amplicon was checked for correct size by electrophoresis on a 1% agarose gel. DNA from clones was purified to remove primer and dNTPs by treatment with exonuclease and shrimp alkaline phosphatase (Amersham, Pittsburg, PA).
Clones were screened by sequencing with primer Y31, 5’-TKACCGCGGCTGCTG-3’ (519–533 reverse). If the clone sequence differed by more than 7 bases from previously identified feline oral reference sequences, the remaining amplicon DNA was purified using QIAquick PCR purification kits (Qiagen, Valencia, CA) and fully sequenced with additional primers (Dewhirst et al., 2012 (link)).

Dewhirst F.E., Klein E.A., Bennett M.L., Croft J.M., Harris S.J, & Marshall-Jones Z.V. (2014). The Feline Oral Microbiome: A Provisional 16S rRNA gene based Taxonomy with Full-length Reference Sequences. Veterinary microbiology, 175(0), 294-303.

+ Open protocol

+ Expand

HER2 Genetic Profiling in Gastric Cancer

Check if the same lab product or an alternative is used in the 5 most similar protocols

The direct sequencing of exon 8, the transmembrane domain (exon 17), and the kinase domain (exons 18‐24) of HER2 was carried out for each of the 12 cell lines and 123 primary gastric cancer samples. The sequences of the primers were the same as previously reported.13, 14 The detailed PCR protocol has been previously reported, and all the PCR products were incubated using exonuclease I and shrimp alkaline phosphatase (Amersham Biosciences). DNA sequence alterations were evaluated using MutationTaster2.15

Yoshioka T., Shien K., Namba K., Torigoe H., Sato H., Tomida S., Yamamoto H., Asano H., Soh J., Tsukuda K., Nagasaka T., Fujiwara T, & Toyooka S. (2018). Antitumor activity of pan‐HER inhibitors in HER2‐positive gastric cancer. Cancer Science, 109(4), 1166-1176.

+ 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?

Revolutionizing how scientists
search and build protocols!