Genomic DNA from wild type and exconjugants from the indicated strains were isolated from liquid cultures using the Blood and Tissue DNeasy kit (Qiagen, Hilden, Germany) after pretreating the cells with 20 mg/mL lysozyme for 0.5–1 h at 30 °C. PCR was performed using control primers beyond the homology regions with KODXtreme Taq polymerase (Millipore, Massachusetts, USA). Where indicated, PCR products were subjected to digest with specific restriction enzymes to differentiate between PCR products of wild type genomic sequences and successful genome editing by knock-ins. Positive samples were purified using ExoSAP-IT™ (Affymetrix USB, Massachusetts, USA) and validated by Sanger sequencing.
Kodxtreme taq polymerase
Manufactured by Merck Group
Sourced in United States
KODXtreme Taq polymerase is a high-fidelity DNA polymerase enzyme produced by Merck Group. It is designed for use in various molecular biology applications that require accurate DNA amplification.
Automatically generated - may contain errors
Lab products found in correlation
5 protocols using kodxtreme taq polymerase
1
Genomic DNA Isolation and PCR Validation
2
Genomic DNA Isolation and Verification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Genomic DNA from wild type and exconjugants from the indicated strains were isolated from liquid cultures using the Blood and Tissue DNeasy kit (Qiagen) after pretreating the cells with 20 mg/mL lysozyme for 0.5–1 h at 30 °C. PCR was performed using control primers beyond the homology regions or knock-in specific primers (Supplementary Table 8 ) with KODXtreme Taq polymerase (Millipore). Where indicated, PCR products were subjected to digest with specific restriction enzymes to differentiate between PCR products of wild type genomic sequences and successful genome editing by knock-ins. Positive samples were purified using Qiaquick PCR purification kit (Qiagen) and validated by Sanger sequencing.
3
Genomic DNA Isolation and Verification
4
Genomic DNA Isolation and PCR Validation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Genomic DNA from wild type and exconjugants from the indicated strains were isolated from liquid cultures using the Blood and Tissue DNeasy kit (Qiagen) after pretreating the cells with 20 mg/mL lysozyme for 30 min at 37 °C. PCR was performed using control primers beyond the homology regions or knock-in specific primers (Additional file 1 : Table S4) with KODXtreme Taq polymerase (Millipore). Where indicated, PCR products were subjected to digest with specific restriction enzymes to differentiate between PCR products of wild type genomic sequences and successful genome editing by knock-ins. Positive samples were validated by Sanger sequencing. Oligos used for PCR and sequencing are listed in Additional file 1 : Table S4.
5
Genomic DNA Extraction and PCR Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Genomic DNA from wild type and exconjugants from the indicated strains were isolated from liquid cultures using the Blood and Tissue DNeasy kit (Qiagen) after pretreating the cells with 20 mg/mL lysozyme for 0.5-1 h at 30 °C. PCR was performed using control primers beyond the homology regions (Supplementary Table 1) with KODXtreme Taq polymerase (Millipore). Where indicated, PCR products were subjected to digest with specific restriction enzymes to differentiate between PCR products of wild type genomic sequences and successful genome editing by knock-ins. Positive samples were purified using ExoSAP-IT TM (Affymetrix USB) and validated by Sanger sequencing (Figure S1,S2).
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
Revolutionizing how scientists
search and build protocols!