long range pcr (New England Biolabs)


Name:
Phusion High Fidelity DNA Polymerase
Description:
Phusion High Fidelity DNA Polymerase 500 units
Catalog Number:
m0530l
Price:
446
Size:
500 units
Category:
Thermostable DNA Polymerases
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Structured Review

Phusion High Fidelity DNA Polymerase 500 units
https://www.bioz.com/result/long range pcr/product/New England Biolabs
Average 99 stars, based on 44 article reviews
Price from $9.99 to $1999.99
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1) Product Images from "Phenotype and function of B cells and dendritic cells from interferon regulatory factor 5-deficient mice with and without a mutation in DOCK2"
Article Title: Phenotype and function of B cells and dendritic cells from interferon regulatory factor 5-deficient mice with and without a mutation in DOCK2
Journal: International Immunology
doi: 10.1093/intimm/dxs114

Figure Legend Snippet: A DOCK2 mutation is detected in IRF5 −/− mice backcrossed 11 generations to C57BL/6 ( IRF5 −/− 11G) mice but not in IRF5 −/− mice backcrossed 15 generations to C57BL/6 ( IRF5 −/− 15G). (A) RT–PCR to detect the DOCK2 mutation (DOCK2mu). RNA was purified from splenic B cells of wild-type C57BL/6 mice from The Jackson Laboratory (Jackson wild-type), IRF5 +/+ and IRF5 −/− 15G littermates and IRF5 −/− ). Primers used in the left-hand gel give a 577-bp product for the DOCK2 mutation and a 383-bp product for wild-type DOCK2. Primers used in the right-hand gel give a 158-bp product for the DOCK2 mutation and do not amplify wild-type DOCK2. (B) The diagram shows the hypothesized differences in genomic DOCK2 DNA between wild-type and DOCK2 mutant IRF5 −/− mice. In the DOCK2 mutant mice, the duplicated exon 28 and 29 together with some flanking DNA is inserted into the intron between exons 27 and 28. The gel shows a PCR performed using a forward primer, which recognizes exon 29 (Ex29F2) and 27 reverse primers (R1-R26 or Ex28R2), which detect the region in the intron between exons 27 and 28 that is closest to exon 28. PCR products were obtained with the R23–R25 primers. IRF5 −/− 11G genomic DNA containing the DOCK2 mutation was used as the template. (C) PCR was performed using the R23 reverse primer and 10 forward primers, which recognize either exon 29 (Ex29F2) or the intron between exons 29 and 30 (In\.1F to In29.9F). (D) DNA sequence of the 3′-end of DOCK2 mutation. The shaded region is the duplicated intronic sequence between exons 29 and 30, and the unshaded region is the non-duplicated intron between exons 27 and 28. (E) Diagram of the DOCK2 mutation. The duplicated segment of the DOCK2 gene present in the DOCK2 mutation ends at 3991bp after exon 29. This duplicated segment is inserted into intron 27–28 at 17 306bp before exon 28. (F) PCR to detect the DOCK2 mutation. Genomic DNA from IRF5 −/− 11G mice gave a PCR product for the DOCK2 mutation (305bp), whereas DNA from IRF5 +/+ and IRF5 −/− 15G littermates did not. CD19 PCR was used as an internal control to verify the adequacy of DNA preparation in each sample.
Techniques Used: Mutagenesis, Mouse Assay, Reverse Transcription Polymerase Chain Reaction, Purification, Polymerase Chain Reaction, Sequencing
2) Product Images from "Structural variation and missense mutation in SBDS associated with Shwachman-Diamond syndrome"
Article Title: Structural variation and missense mutation in SBDS associated with Shwachman-Diamond syndrome
Journal: BMC Medical Genetics
doi: 10.1186/1471-2350-15-64

Figure Legend Snippet: Additional studies of the genomic segment that includes SBDS supports the presence of an insertion of unknown origin within the paternal allele. a . Agarose gel analysis of a 13.1 kb long-range PCR product spanning SBDS and flanking region, the region targeted by the Southern blotting studies (Figures 2 and 3 ). DNA from patient (BAB3762), parents (BAB3763- mother, BAB3764-father) and a normal controls (CTL1 and CTL2) were amplified using primers DelFb + KpnR, and then digested with either Kpn I or Sac I. Kpn I did not digest the 13.1 kb PCR product, consistent with lack of amplification of the paternal allele. Consistently, Sac I digestion of the 13.1 kb PCR product showed an identical pattern in samples and controls. b . Sanger sequencing of intron 2 amplified along with exon 2 using short range PCR revealed inconsistent segregation of a paternal genotype for two polymorphic SNPs in the patient (BAB3762). *Non-digested PCR product.
Techniques Used: Agarose Gel Electrophoresis, Polymerase Chain Reaction, Southern Blot, Amplification, Sequencing
3) Product Images from "A one-tube method for rapid and reliable plant genomic DNA isolation for PCR analysis"
Article Title: A one-tube method for rapid and reliable plant genomic DNA isolation for PCR analysis
Journal: bioRxiv
doi: 10.1101/2020.02.13.948455

Figure Legend Snippet: DNA extracts using the one-tube protocol from various Arabidopsis tissues and leaf biomass have negligible inhibitory effect on PCR analysis. For leaf biomass and size comparison, 1-2 mg is approximately equivalent to one cotyledon; 5 mg is equivalent to the leaf mass excised using the inner cap of a 1.5 ml microcentrifuge tube; 25-50 mg is equivalent to one single fully developed rosette leaf.
Techniques Used: Polymerase Chain Reaction

Figure Legend Snippet: PCR analysis of DNA extracts prepared using the standard Edwards protocol (1), the one-tube protocol (2 and 3) and the one-step protocol of Kasajima et al (4). Two independent DNA extracts from (2) to (4) are shown; DNA extracts of (3) were stored at room temperature for three months.
Techniques Used: Polymerase Chain Reaction

Figure Legend Snippet: DNA extracted by the one-tube protocol is effectively amplified by long-range PCR using LongAmp Taq DNA polymerase. Two independent DNA extracts using Edwards (ED) and one-tube (OT) protocols are shown.
Techniques Used: Amplification, Polymerase Chain Reaction
4) Product Images from "piggyBac as a high-capacity transgenesis and gene-therapy vector in human cells and mice"
Article Title: piggyBac as a high-capacity transgenesis and gene-therapy vector in human cells and mice
Journal: Disease Models & Mechanisms
doi: 10.1242/dmm.010827
![PB -mediated BAC transgene expression in mice. (A) RT-PCR analysis showed RORγ but not RORγt expression in ... PB -mediated BAC transgene expression in mice. (A) RT-PCR analysis showed RORγ but not RORγt expression in the liver, kidney and muscle of wild-type (WT) and transgenic mice. (B) Quantitative real-time PCR with tail genomic DNA revealed different copy numbers of PB[BAC] in each of the the transgenic lines. (C) Real-time RT-PCR showed elevated RORγt expression in the thymus of transgenic animals. (D,E) Similar levels of increased RORγ expression were also observed in the kidney (D) and liver (E). All samples were taken from 20-day-old mice. Line numbers are indicated. *** P](https://storage.googleapis.com/bioz_article_images/PMC3634665/DMM010827F4.jpg)
Figure Legend Snippet: PB -mediated BAC transgene expression in mice. (A) RT-PCR analysis showed RORγ but not RORγt expression in the liver, kidney and muscle of wild-type (WT) and transgenic mice. (B) Quantitative real-time PCR with tail genomic DNA revealed different copy numbers of PB[BAC] in each of the the transgenic lines. (C) Real-time RT-PCR showed elevated RORγt expression in the thymus of transgenic animals. (D,E) Similar levels of increased RORγ expression were also observed in the kidney (D) and liver (E). All samples were taken from 20-day-old mice. Line numbers are indicated. *** P
Techniques Used: BAC Assay, Expressing, Mouse Assay, Reverse Transcription Polymerase Chain Reaction, Transgenic Assay, Real-time Polymerase Chain Reaction, Quantitative RT-PCR
![... PB[BAC] insertions in human 293 cells. (A–F) Inverse PCR with primers PBLinvB1 and PBLinvF1 (A) recovered single ... Molecular analysis of PB[BAC] insertions in human 293 cells. (A–F) Inverse PCR with primers PBLinvB1 and PBLinvF1 (A) recovered single insertions in most of the clones (B). The insertion sites were confirmed by genotyping PCR using pairs of primers targeting both flanking genomic sequences (G-RL or G-RB) and PB terminal sequences (PBL-B or PBR-F), as shown in panel C. Integrity of PB[BAC] transgenes was analyzed by PCR using ten primer pairs evenly spaced along the BAC insert (fragments 1–10 in panel D). Two clones showed all positive results (E) and were further examined by long-range PCR using 21 pairs of overlapping primers covering the full length (fragments A–U in panel D), as shown in panel F and supplementary material Fig. S2 . M, 1 kb ladder; +, positive control with PB[BAC] as the template, −, negative control with 293 genomic DNA as the template; stars mark location of SNPs used to verify BAC integrity in transgenic mice.](https://storage.googleapis.com/bioz_article_images/PMC3634665/DMM010827F2.jpg)
Figure Legend Snippet: Molecular analysis of PB[BAC] insertions in human 293 cells. (A–F) Inverse PCR with primers PBLinvB1 and PBLinvF1 (A) recovered single insertions in most of the clones (B). The insertion sites were confirmed by genotyping PCR using pairs of primers targeting both flanking genomic sequences (G-RL or G-RB) and PB terminal sequences (PBL-B or PBR-F), as shown in panel C. Integrity of PB[BAC] transgenes was analyzed by PCR using ten primer pairs evenly spaced along the BAC insert (fragments 1–10 in panel D). Two clones showed all positive results (E) and were further examined by long-range PCR using 21 pairs of overlapping primers covering the full length (fragments A–U in panel D), as shown in panel F and supplementary material Fig. S2 . M, 1 kb ladder; +, positive control with PB[BAC] as the template, −, negative control with 293 genomic DNA as the template; stars mark location of SNPs used to verify BAC integrity in transgenic mice.
Techniques Used: BAC Assay, Inverse PCR, Polymerase Chain Reaction, Genomic Sequencing, Clone Assay, Positive Control, Negative Control, Transgenic Assay, Mouse Assay
5) Product Images from "Removal of Integrated Hepatitis B Virus DNA Using CRISPR-Cas9"
Article Title: Removal of Integrated Hepatitis B Virus DNA Using CRISPR-Cas9
Journal: Frontiers in Cellular and Infection Microbiology
doi: 10.3389/fcimb.2017.00091

Figure Legend Snippet: Analysis of integrated HBV DNA in the stable HBV cell line A64. (A) Integrated HBV DNA in the stable HBV A64 cell line and the gRNA target sites in the repeat region of the 1.1 HBV genome copy. (B) The 4,049-bp DNA fragment representing the 3,362-bp integrated HBV DNA (1.1 copies) plus a 687-bp pTriexHBV1.1-derived flanking sequence was efficiently amplified from cellular genomic DNA using the integrated HBV-specific primers P1 and P2. (C) PCR analysis using the A1AT and HBV S-gene primer sets conducted on total genomic DNA and circular duplex DNA to assess the effect of extraction on circular duplex DNA. (D) PCR analysis of the integrated HBV DNA or circular duplex DNA isolated from cells using the P1/P3 and S-gene primer sets. Using P1 and the HBV core region-specific primer P3, the HBV S-gene amplicons were predicted to be 542- and 572-bp, respectively. Primers P1/P3 did not amplify circular duplex DNA.
Techniques Used: Derivative Assay, Sequencing, Amplification, Polymerase Chain Reaction, Isolation

Figure Legend Snippet: CRISPR-Cas9/gRNA-69 efficiently removed the integrated HBV genome from a stable HBV cell line. (A) Analysis of PCR amplicon lengths using a primer pair (P1 and P2) targeting the integrated HBV-flanking sequence revealed elimination of the full-length integrated HBV genome (3,173-bp), leaving one fragment (873-bp predicted segment from its flanking region). (B) Diagram showing excision of the full-length integrated HBV genome. The remaining fragment included the expected 687-bp from the integrated HBV flanking sequence and a 186-bp HBV repeat core region sequence. (C) Sanger sequencing of the remaining fragment (873-bp) showing the HBV flanking sequence (small letters, 687-bp) and the partial sequences (189 − 3 = 186-bp) of the integrated HBV repeat region B (green) and repeat region A (red) with a 3-bp deletion around the gRNA-69 targeting site (yellow-highlighted). Elimination of the full-length integrated HBV genome is indicated by a strikethrough. (D,E) The amounts of HBeAg, HBsAg and HBV DNA in cell culture supernatants and HBV cccDNA in the gRNA-empty-treated group (K-15) and gRNA-69-treated group (69-7) over 300 consecutive days. The HBsAg and HBeAg test results in the gRNA-69-treated group (69-7) were always under the negative threshold (0.05 IU/ml for HBsAg and 1 S/CO for HBeAg), and the amounts of HBV DNA and HBV cccDNA in the supernatants were always undetectable (
Techniques Used: CRISPR, Polymerase Chain Reaction, Amplification, Sequencing, Cell Culture
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