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Phase lock heavy tubes

Manufactured by Quantabio
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Phase Lock heavy tubes are designed for use in DNA extraction and sample preparation workflows. They feature a specialized gel-based barrier that facilitates the separation of aqueous and organic phases during sample processing. These tubes are constructed with high-quality materials to withstand centrifugation and other laboratory procedures.

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Lab products found in correlation

Rtcb ligase, by New England Biolabs (2 mentions) T4 polynucleotide kinase, by New England Biolabs (2 mentions) Rnaeasy kit, by Qiagen (1 mentions) Qiashredder, by Qiagen (1 mentions) Sodium dodecyl sulfate (sds), by Thermo Fisher Scientific (1 mentions) Eb buffer, by Qiagen (1 mentions) Proteinase k, by Merck Group (1 mentions)

Close spelling variants of this product

Heavy Phase Lock Gel tubes Phase Lock Gel Heavy tubes 5PRIME Phase Lock Gel Heavy tubes

5 protocols using phase lock heavy tubes

1

RNA Purification and Ligation

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After gel purification of autocatalytically cleaved RNA, 300 pmol were treated with T4 polynucleotide kinase (New England Biolabs M0201) according to the manufacturer’s protocol at 37 °C for 30 min and inactivated for 20 min at 65 °C. The products were cleaned by phenol chloroform extraction using heavy phase-lock tubes (Quantabio 2302830). 10 pmol of this purified T4-PNK-treated RNA or of the gel purified RNA was ligated using RtcB Ligase (New England Biolabs M0458) for 1 h at 37 °C.
Litke J.L, & Jaffrey S.R. (2019). Highly efficient expression of circular RNA aptamers in cells using autocatalytic transcripts. Nature biotechnology, 37(6), 667-675.
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2

RNA Purification and Ligation

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After gel purification of autocatalytically cleaved RNA, 300 pmol were treated with T4 polynucleotide kinase (New England Biolabs M0201) according to the manufacturer’s protocol at 37 °C for 30 min and inactivated for 20 min at 65 °C. The products were cleaned by phenol chloroform extraction using heavy phase-lock tubes (Quantabio 2302830). 10 pmol of this purified T4-PNK-treated RNA or of the gel purified RNA was ligated using RtcB Ligase (New England Biolabs M0458) for 1 h at 37 °C.
Litke J.L, & Jaffrey S.R. (2019). Highly efficient expression of circular RNA aptamers in cells using autocatalytic transcripts. Nature biotechnology, 37(6), 667-675.
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3

RNA Extraction from Cells and Extracellular Vesicles

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Total RNA was isolated from 100K EV pellets and cells using a combination of the RNAeasy kit (Qiagen) and Phase Lock Heavy Tubes (Quanta Bio). Briefly, ~ 6 × 106 cells or 10–50 μg of EVs were lysed with 750 μL Qiazol after which 350 μL of chloroform was added and mixed by pipette. The slurry was transferred to a phase lock tube for phase separation. The resulting aqueous phase containing the RNA was processed according to the Qiagen protocol and purified RNA was eluted in either 50 μL (cells) or 30 μL (EVs).
Zhang P., Crow J., Lella D., Zhou X., Samuel G., Godwin A.K, & Zeng Y. (2018). Ultrasensitive Quantification of Tumor mRNAs in Extracellular Vesicles with an Integrated Microfluidic Digital Analysis Chip. Lab on a chip, 18(24), 3790-3801.
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4

DNA Extraction from Cells

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FACS-enriched or baseline cells were collected by centrifugation, washed with PBS and a maximum of 5E6 cells were resuspended in 350 µL DNA extraction buffer (10 mM Tris-HCl, 150 mM NaCl, 10 m M EDTA, pH 8.0). After addition of each 3.5 µL 10% SDS (Invitrogen) and 20 mg/mL proteinase K (Sigma), the suspension was incubated at 55 °C overnight. proteinase K was heat inactivated at 95 °C for 10 min, followed by incubation with 10 mg/mL RNase A at 37 °C for 30 min. The mixture was transferred to a QIAshredder (Qiagen) and spun at 20,000 g for 2 min. Phase Lock heavy tubes (Quantabio) were spun and 350 µL phenol/chloroform/isoamyl alcohol (Roth) was added. Samples were mixed by shaking. Samples were spun at 20,000 g for 8 min. The upper phase was chloroform extracted twice with 350 µL chloroform, and DNA was ethanol-precipitated (35 µL 3 M Na-acetate pH 5.2 and 1050 µL absolute ethanol) at − 20 °C overnight. Samples were centrifuged at 20,000×g for 30 min. The DNA pellet was washed twice with 70% ethanol, dried and resuspended in 50 µL buffer EB (Qiagen).
Lindner B., Martin E., Steininger M., Bundalo A., Lenter M., Zuber J, & Schuler M. (2021). A genome-wide CRISPR/Cas9 screen to identify phagocytosis modulators in monocytic THP-1 cells. Scientific Reports, 11, 12973.
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5

Extraction and Purification of Genomic DNA

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To extract DNA, cells were centrifuged at ~ 800 g in a tabletop IEC centrifuge for 3 minutes and cell pellets (~ 1 × 107 cells) were resuspended in ~200 μl of supernatant, distributed equally to two 2.2ml Eppendorf tubes containing 300 μl of 55°C NDS Proteinase K solution, freshly made by combining 2 parts NDS (1% SDS, 10 mM Tris, pH 8.0, 0.5 M EDTA, pH 8.0) and 1 part Proteinase K (2 mg/ml in H2O), and incubated at 55°C overnight without RNase. Cell extracts were diluted with 400 μl of low EDTA TE (10 mM Tris, pH 8.0; 0.1 mM EDTA) and extracted with 800 μl of Phenol:Chloroform:Isoamyl Alcohol (25:24:1 v/v) and 800 μl Chloroform:Isoamyl Alcohol (25:24:1 v/v) in Phase Lock heavy tubes (QuantaBio). DNA was precipitated with 1 ml cold 100% ethanol, washed with 500 μl cold 70% ethanol and 500 μl cold 100% ethanol, and air dried. DNA was resuspended in 400 μl low EDTA TE buffer (PH 8.0). For PCR, total DNA was adjusted to 100 ng/μl and 1μl used for each PCR reaction.
Cassidy-Hanley D.M., Doerder F.P., Hossain M., Devine C, & Clark T. (2022). Molecular identification of Tetrahymena species. The Journal of eukaryotic microbiology, 70(1), e12936.
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