Gel recovery kit

Manufactured by Solarbio

Sourced in China

The Gel Recovery Kit is a tool for extracting and purifying DNA fragments from agarose gels. It provides a simple and efficient method to isolate and recover DNA of interest from gel electrophoresis. The kit includes the necessary solutions and columns to facilitate the DNA extraction process.

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Lab products found in correlation

Vortex mixer, by Scilogex (1 mentions) Dna extraction kit, by Tsingke (1 mentions) Dh5α competent cells, by Thermo Fisher Scientific (1 mentions) Lipofectamine 3000 transfection reagent, by Thermo Fisher Scientific (1 mentions) Reverse transcription kit, by Solarbio (1 mentions) Total rna extraction kit, by Solarbio (1 mentions)

3 protocols using gel recovery kit

Isolation and Identification of Calcium-Solubilizing Bacteria

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The collected samples were mixed with a vortex mixer (Scilogex, United States) for 5 min, serially diluted (10⁻¹ to 10⁻⁹), and 0.1 mL of each of the 10⁻³ to 10⁻⁷ dilutions was applied to MRS agar containing calcium carbonate. Individual colonies with soluble calcium circles were selected for passaging on MRS agar and incubated for 48 h at 37°C. After three passages, the isolates were stored frozen in 50% (w/v) sterile glycerol at −80°C. Supplementary Figure S1 shows the morphological and gram staining results of GLP02 and GLP06.
To identify the isolates, genomic DNA was extracted using a DNA extraction kit (Tsingke, China). Polymerase chain reaction (PCR) amplification was performed using the primers 16S-27F (5’-AGAGTTTGATCCTGGCTCAG-3′) and 16S-1492R (5’-TACGGCTACCTTGTTACGACTT-3′) under the following conditions: pre-denaturation at 94°C for 5 min, denaturation at 94°C for 45 s, annealing at 55°C for 30 s, and extension at 72°C for 45 s for 30 cycles, then extension at 72°C for 10 min. The PCR products were identified by 1.0% agarose gel electrophoresis, and the target bands were recovered with a gel recovery kit (Solarbio, China) and sent to Tsingke Biological Technology for sequencing. A phylogenetic tree was constructed using Mega 11.0 software to determine the evolutionary relationships of the isolated strains.

Zhao M., Liu K., Zhang Y., Li Y., Zhou N, & Li G. (2023). Probiotic characteristics and whole-genome sequence analysis of Pediococcus acidilactici isolated from the feces of adult beagles. Frontiers in Microbiology, 14, 1179953.

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Cloning tLyp-1-lamp2b Fusion Protein

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Total RNA was extracted from Hela cells using total RNA extraction kit, which was reversely transcribed into cDNA using reverse transcription kit (Solarbio, Beijing, China). tLyp-1 +  lamp2b gene was amplified by PCR using cDNA as template, purified and recovered by using gel recovery kit (Solarbio, Beijing, China). The pEGFP-C1 and tLyp-1-lamp2b genes were digested by Bgl II and BamH I enzymes, then connected by T4 ligase. The connection products were transformed into DH5α competent cells (stored at −80 °C, Thermo Scientific, Waltham, USA) and cultured for 12 h. Single colony was culled for plasmid extraction. Lipofectamine 3000 transfection reagent (Invitrogen, USA) were used to transfect pEGFP-C1 plasmid vector expressing tLyp-1-lamp2b fusion proteins into HEK293T cells.

Bai J., Duan J., Liu R., Du Y., Luo Q., Cui Y., Su Z., Xu J., Xie Y, & Lu W. (2019). Engineered targeting tLyp-1 exosomes as gene therapy vectors for efficient delivery of siRNA into lung cancer cells. Asian Journal of Pharmaceutical Sciences, 15(4), 461-471.

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Lentiviral Overexpression of LAMP-2B and HIF1α Knockdown

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The LAMP-2B + 5′-TGTTGTGGTAAACGTAAA-3′ (gene sequence of CCGKRK) gene was amplified from cDNA template by PCR, then purified and recovered with a gel recovery kit (Beijing Solarbio Science & Technology Co., Ltd.). The gene was subsequently cloned and ligated to the GV493 vector (hU6-MCS-CBh-MCS-IRES-puromycin; Shanghai GeneChem Co., Ltd.). A 3rd generation system was used to package the lentivirus. Additionally, an sh-HIF1α sequence (5′-ACGACAAGAAAAAGATAAGTT-3′) was also ligated into the same GV493 vector using an independent promoter. Upon transduction into cells, the lentiviral particles generated in this manner facilitated the overexpression of both LAMP-2B + CGKRK and HIF1α shRNA. The vectors (100 nM) and packaging plasmids (vector:packaging vector:envelope ratio, 10:3:1) were co-transfected into 1×10⁶ 293T cells with Lipofectamine 2000 at 37°C for 8 h. The sh-HIF1α-LAMP-2B lentivirus and negative control (empty vector) were collected and filtered through a 0.45 µM filter after 293T cells were cultured at 37°C for 3 days. MSCs were then transduced with lentiviral vectors at a MOI of 10 for 8 h followed by replacement with fresh medium. MSCs were cultured at 37°C for 48 h and subsequently treated with puromycin (1 µg/ml) for 48 h to select stably transduced cells, 0.5 µg/ml puromycin was used for maintenance.

Chen F., Cen H., Mao D, & Feng R. (2023). Placental homing peptide guides HIF1α‑silenced exosomes conjugates for targeted enhancement of invasion of trophoblast cells. Molecular Medicine Reports, 28(1), 135.

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