The following antibodies were used throughout this study: from Abclonal (Wuhan, China), anti-FLAG (AE063), anti-HA (AE036), anti-ASGR1 (A13279), anti-β-Actin (AC026). From Abcam (Cambridge, UK), anti-ASGR1 (ab254262). From proteintech (Wuhan, China), anti-ACE2 (21115-1-AP). From Sino Biological (Beijing, China), rabbit-anti-SARS-CoV-2 NP antibody. From R&D systems (UMN, USA), anti-His-Alexa Fluor488-conjugated Antibody (IC050G). Anti-Flag Magnetic Beads (HY-K0207) and ACE2/ANG (1-7) inhibitor-A779 (HY-P0216) were purchased from MCE. The Sipke of SARS-CoV-2 (DRA59) was purchased from novoprotein (Shanghai, China). 2× Taq Master Mix (P112) and High-fidelity PCR enzyme-2× Phanta Max Master Mix (P515) were purchased from Vazyme (Nanjing, China). PMD18-T (6011) was purchased from Takara (Beijing, China). Cell Genome Extraction Kit (DP304) and Plasmid Extraction Kit (DP103, DP108, DP117) were purchased from Tiangen (Beijing, China). Gel Extraction Kit (CW2302) was purchased from CWBIO (Nanjing, China). Luciferase detection kit (E6110) was purchased from Promega (Madison, USA). Cell Counting Kit (CCK-8) and TUNEL Apoptosis Detection Kit (FITC) were purchased from Yeasen (Shanghai, China).
Cell genome extraction kit
Manufactured by Tiangen Biotech
Sourced in China
The Cell Genome Extraction Kit is a laboratory tool designed to isolate and extract genomic DNA from various cell types. The kit provides a standardized protocol and reagents necessary for the effective separation and purification of DNA samples, enabling downstream analyses and applications.
Automatically generated - may contain errors
Lab products found in correlation
Plasmid extraction kit, by Tiangen Biotech (2 mentions)
Gel extraction kit, by CWBIO (2 mentions)
High fidelity pcr enzyme 2 phanta max master mix, by Vazyme (2 mentions)
Cell counting kit cck 8, by Yeasen (2 mentions)
Luciferase detection kit, by Promega (1 mentions)
Tunel apoptosis detection kit, by Yeasen (1 mentions)
Rabbit anti sars cov 2 np antibody, by Sino Biological (1 mentions)
Pmd18 t 6011, by Takara Bio (1 mentions)
La pcr buffer 2, by Takara Bio (1 mentions)
Taq master mix, by Vazyme (1 mentions)
Pmd18 t, by Takara Bio (1 mentions)
Anti hdac1, by Abcam (1 mentions)
Anti hdac2, by Abcam (1 mentions)
Anti β actin, by Cell Signaling Technology (1 mentions)
Recombinant human ifn γ, by R&D Systems (1 mentions)
Anti h3k18ac, by Abcam (1 mentions)
Recombinant human ifn α, by R&D Systems (1 mentions)
Anti yy1, by Cell Signaling Technology (1 mentions)
4 protocols using cell genome extraction kit
1
Antibody Sources for SARS-CoV-2 Research
2
CRISPR-mediated Editing of EDAR Gene
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Wild-type Alba white Cashmere GFbs were obtained and used as controls. These GFbs were transfected by electroporation, either with the sgRNA1 and Cas9 plasmids, or the sgRNA2 and Cas9 plasmids. At 48 h post-transfection, the gDNAs of the modified and wild-type cells were extracted using the cell genome extraction kit (TianGen Biotech, Beijing, China). These gDNAs were then used as templates to amplify the target sequence of the EDAR gene via PCR with the following primers 5′-GTGGTGGTCGTCGTGGTGATGC-3′ (forward) and 5′-CTGCTCAGCCTTCCTTATGGTC-3′ (reverse). The PCR conditions used were as described above. The resulting amplicons were purified and mixed with 10× La PCR Buffer II (TaKaRa Bio, Shiga, Japan), and a heteroduplex was formed by gradient annealing under the following conditions: 95°C for 10 min, 95 to 85°C ramping at -2°C/s, 85 to 25°C at -0.3°C/s, and a 4°C hold. The heteroduplexes were processed using the Surveyor Mutation Detection Kit (Transgenomic, Omaha, NE, USA) and run on a 2% agarose gel to detect mutation efficiency.
3
Antibody Characterization and Validation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following antibodies were used throughout this study: anti-FKBP3 from Thermo Fisher (catalog no. PA5-19483); anti-HDAC1 (catalog no. ab280198), anti-HDAC2 (catalog no. ab219053), anti-H3K4ac (catalog no. ab176799), and anti-H3K18ac (catalog no. ab40888) from Abcam (Cambridge, UK); and anti-YY1 (catalog no. 46395) and anti-β-actin (catalog no. 4970) from Cell Signaling Technology (MA, USA). We purchased 2× Taq master mix (catalog no. P112) and high-fidelity PCR enzyme 2× Phanta Max master mix (catalog no. P515) from Vazyme (Nanjing, China). PMD18-T (catalog no. 6011) was purchased from TaKaRa (Beijing, China). Cell genome extraction kit (catalog no. DP304) and plasmid extraction kit (catalog nos. DP103, DP108, and DP117) were purchased from Tiangen (Beijing, China). Gel extraction kit (catalog no. CW2302) was purchased from CWBio (Nanjing, China). Luciferase and NanoLuc detection kits (catalog nos. E6110 and N1110, respectively) were purchased from Promega (Madison, USA). Recombinant human IFN-α (catalog no. 11200-1), recombinant human IFN-β (catalog no. 8499-IF), and recombinant human IFN-γ (catalog no. 285-IF) were purchased from R&D Systems (USA). Cell counting kit (CCK-8) and TUNEL apoptosis detection kit (fluorescein isothiocyanate [FITC]) were purchased from Yeasen (Shanghai, China).
4
Nanopore Sequencing for Genome Editing
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cells were trypsinized and collected, and genomic DNA was extracted using the Cell Genome Extraction kit (Tiangen Biotech, Beijing, China) for Oxford Nanopore sequencing library construction. High-throughput sequencing was then performed on MinION. Porechop (version 0.2.4) was used to remove adapter sequences from Nanopore reads. Minimap2 (version 2.17-r941) was utilized to map long reads to the human genome. Samtools (version 1.9) was employed to process bam files. bamCoverage (version 3.3.0) was used to convert bam files to bigwig files. Continuous genome reference sequences of three different strategies were also established by predicting the precise connection after editing, and then Nanopore reads were mapped to this reference sequence to explore change models and reads coverage around the cut sites. Reads coverage in bigwig files or bam files was visualized using the UCSC genome browser or the IGV genome browser. Density plot and mutation percentage of bases were counted by custom R scripts.
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!