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Bio-protocol Preprint Repository
A preprint repository for life science protocols
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Monoclonal phage ELISA
Q&A Published: Nov 5, 2026Monoclonal phage ELISAAfter the final round of screening, add 50 µL of OmniMAX bacteria (OD600 ~0.6) to each well and incubate at room temperature for 20 min with shaking at 600 rpm to allow phage infection.Collect all the bacteria. Take 100 µL bacteria for 10-fold serial dilutions, spread the diluted bacteria on ampicillin LB plates (without helper phage infection), and incubate the plates at 37 °C overnight.The next day, add 100 µL of 2×YT medium supplemented with ampicillin to each well of a ...
Ferric reducing antioxidant potential (FRAP) assay
Q&A Published: Nov 5, 2026Protein hydrolysates derived from residual after polysaccharide extraction of Chlorella vulgaris biomass improves yield and quality of baby leaf lettuceAnnamaria Di Serio1#, Giovanna Aquino2,3#, Francesco del Prete2, Francesca Sansone2,5, Emanuela Salviati2, Manuela Giovanna Basilicata3, Michele Manfra4, Pietro Campiglia2, Domenico Ronga2,* and Giacomo Pepe2 1 Chemistry and Biology Department, University of Salerno, Via Giovanni Paolo II 132, Fisciano, 84084, Italy. adiserio@unisa.it2 Departmen...
Osteogenic differentiation and staining of BMSCs
Q&A Published: Nov 5, 2026Osteogenic differentiation and staining of BMSCs Authors Weizhou Wang wangweizhou@kmmu.edu.cn Fei He drhefei@126.comShengyu Long 1043873657@qq.comAbstractBone marrow mesenchymal stem cells(BMSCs) possess multilineage differentiation potential and can differentiate into osteoblasts under the influence of specific biofactors. The quantification of osteoblast-specific products can be achieved through alkaline phosphatase (ALP) and Alizarin Red S (ARS) staining, th...
Size-Exclusion Chromatography of Argonaute-Containing Complexes
Q&A Published: Nov 4, 2026Size-Exclusion Chromatography for the Analysis of Argonaute-Containing Ribonucleoprotein Complexes in Mammalian Cells and TissuesIntroductionThis protocol describes size-exclusion chromatography (SEC) to separate Argonaute (AGO)-containing ribonucleoprotein complexes from mammalian cells and tissues according to apparent molecular size. Clarified native extracts are fractionated on a Superose 6 column; proteins from individual fractions are recovered by trichloroacetic acid (TCA) precipitation a...
Low-cost Medium-Throughput Protein Expression and Purification in E. coli
Q&A Published: Oct 29, 2026AbstractAs the cost of synthetic gene synthesis has decreased, more laboratories have access to large libraries of synthetic genes from which to express and purify proteins. However, current protocols for protein expression and purification are either too slow and laborious to work at large-scale, or require specialist equipment, such as robotic liquid handlers. Since many researchers work at a level between these extremes, we present a medium-throughput protocol to express and purify libraries ...
MicroC library preparation from Drosophila embryos
Q&A Published: Oct 27, 2026Abstract Micro-C maps chromatin contacts by digesting crosslinked chromatin with micrococcal nuclease (MNase) and ligating DNA ends that are held in proximity. This protocol describes library preparation from Drosophila melanogaster embryos, including embryo collection, sequential crosslinking, nuclei isolation, MNase digestion, end processing and biotin labeling, proximity ligation, removal of biotin from unligated ends, DNA purification, and sequencing library preparation. Biotinylated DNA is ...
Meiotic chromosome surface spreads
Q&A Published: Oct 23, 2026Preparation of Mouse Fetal Ovary spread for cytological analysis1. Collection of Embryonic OvariesC57BL/6J female mice were mated with male mice overnight. The following morning, females were examined for the presence of vaginal plugs. The day on which a vaginal plug was detected was designated embryonic day (E) 0.5. At E17.5, pregnant females were euthanized in accordance with the protocol approved by the relevant Ethics Committee. The uterus was removed, and the embryos were dissected and tran...
Construction of Pf-5 mutants and derivative strains
Q&A Published: Oct 21, 2026Protocol for making a deletion of an interested gene in the chromosome of strain Pf-5Day 1: Streak mutant Pf-5 strain on KB plate (no antibiotics) and grow overnight at 28°C.Streak S17-1 E. coli strain carrying deletion construct carried by pEX18Tc to be introduced on LB + Tc20 plate and grow overnight at 37°C.Day 2: Start 4 mL KB culture of Pf-5 strain and shake overnight at 28°C. Start a 4 mL LB + Tc10 culture of S17-1 strain and shake overnight at 37°C. (Note: The S17-1 strains tend to grow ...
In vivo model of liver metastasis by splenic injection.
Q&A Published: Oct 21, 2026Protocol for Establishing a Mouse Model of Colorectal Cancer Liver Metastasis by Intrasplenic Injection of MC38 Cells Followed by Splenectomy1. ObjectiveTo establish an experimental model of colorectal cancer liver metastasis (CRLM) by injecting MC38 murine colorectal cancer cells into the spleen of C57BL/6 mice, thereby introducing tumor cells into the liver through the portal circulation. Subsequent splenectomy is performed to minimize tumor growth within the spleen.This model is used to inves...
Single-molecule motility assays and TIRF microscopy imaging
Q&A Published: Oct 20, 2026Pre-made stock solutions 5XDLB with no KOAc150mM HEPES pH7.4 with NaOH10mM MgOAc 5mM EGTA50% glycerol 5X TEV50mM Tris.HCl pH8 750mM KCl50% glycerol 5X GF150 Buffer125mM HEPES pH7.4750mM KCl5mM MgCl2 300 nM Lis1 WT, del 298-308. Made MTs 488 LAST WEEK ProteinConc. Dim.Bufferdynein TMR from Aga K made on 01/27/2022- (A3-6) clear tubes197 nMGF150 buffer: 25mM HEPES pH7.4, 150mM KCl, 1mM MgCl2, 1mM DTT, and 10% glycerolDynactin unlabeledFrom 10/06/20 - Andrea's rack (B7-5)190 nMGF150 buffer: 2...