- Home
- Protocols
-
Bio-protocol Preprint Repository
A preprint repository for life science protocols
Request a Protocol (RaP)
Request a detailed protocol from the "Materials and Methods" section of any research article.
Fusion inhibition assay using split GFP
Q&A Published: Sep 2, 2026Split GFP cell-cell fusion assayDay 1: seeding HEK293T cellsIn 96 well plate: cells will be transfected with GFP11 and will not be detachedIn 6 well plate: cells will be transfected with Spike + GFP1-10 and will be detached + added to cells in 96 well plate- Cell seeding in 96 well plates: 8000 cells/well, to transfect the next day. If transfecting on day 3: seed 4000 cells/well- Cell seeding in 6 well plates: 200 000 cells/well, to transfect the next day. If transfecti...
Leaf Rolling, Leaf Drying, and Drought Recovery Scores
Q&A Published: Sep 1, 2026Field scoring of rice leaf rolling, leaf drying and recovery under reproductive-stage droughtMethod used in the rainout-shelter experiments reported by Ghneim-Herrera et al. (2017), with visual scores based on the IRRI Standard Evaluation System for Rice (SES). ScopeThis protocol describes the field procedure used to follow leaf rolling (LR), leaf drying (LD) and drought recovery (DRR) in rice exposed to water deficit at the reproductive stage. The observations are non-destructive and are made o...
RiboTag Immunoprecipitation Protocol
Q&A Published: Aug 31, 2026Pierce™ Protein A/G Magnetic Beads Thermo Scientific 88802 Prepare 2x CHX stock, 5mg/ml for washing intestine Anti-HA (abcam ab9110) Homogenization buffer (HB) AFinal Concentration Stock 10 ml Location1NP-401%10%1 mlChemical cabinet2KCl0.1M2M500 ul 3TRIS pH 7.450mM1M500 ulChemical cabinet4MgCl2120mM1M120 ulLibrary room5water 7800 ul Supplemented homogenization buffer (HB-S) AFinal Concentration StockFinal Volume (1.5ml) Location1Homonogenization buffer(HB) 1.40 ml 2Cyclohexamide (Protein inhibit...
Lab protocol to quantify siRNA strands via stem loop qPCR
Q&A Published: Aug 28, 2026Declaimer: The protocol has been optimized to work with snap frozen or RNAlater preserved liver and kidney samples from unchallenged C57BL/6 mice. However, this protocol can be used as a starting point to expand to other tissues. Material used:Lysis buffer (QuantiGene, QuantiGene™ Sample Processing Kit, tissues (Cat. No. QS0106, Lot: 378856-000)+ Proteinase K (50µg/µl, from QuantiGene Kit above)Tissue Lyser (Retch Type MM300)Tungsten Carbide Beads (Qiagen Ref: 69997; Qiagen, Venlo, Netherlands)...
HaCaT cell culture
Q&A Published: Aug 27, 2026Protocol for Culturing HaCaT Cells Materials and ReagentsHaCaT cellsDMEM containing 4.5 g/L glucose, sodium pyruvate, and GlutaMAXFetal bovine serum (FBS)Penicillin-streptomycinPhosphate-buffered saline (PBS)AccutaseDMSOCell counter or hemocytometerComplete Culture MediumPrepare DMEM containing 4.5 g/L glucose, sodium pyruvate, and GlutaMAX, supplemented with 10% FBS and 1% penicillin-streptomycin. Warm the required volume before use and handle all reagents aseptically.Culture ConditionsIncubate...
Quantification of DNA strand breaks by TUNEL assay in Mycobacterium tuberculosis
Q&A Published: Aug 26, 2026AbstractThe primary niche of Mycobacterium tuberculosis (Mtb) is inside phagocytic cells, where the bacteria are exposed to reactive oxygen species (ROS) and reactive nitrogen intermediates (RNIs). [1], [2] The ROS and RNIs can introduce single-stranded or double-stranded DNA breaks in bacteria. This protocol describes how to quantify the DNA strand breaks in Mtb. Materials and ReagentsIn Situ Cell Death Detection Kit, TMR red (Roche Molecular Biochemicals, Indianapolis, IN, Cat. No.- 12156792...
Isolation of microglia and border-associated macrophages
Q&A Published: Aug 19, 2026Mice were decapitated, brains were harvested and placed on ice-cold 2% FBS Iscove's Modified Dulbecco's Medium (IMDM).Cut the large brain into small pieces. Then digest brain tissue in 5mL 2% FBS IMDM with 2U/mL DNase Ⅰ (Roche), 1 mg/mL Collagenase D (Roche) and 2U/mL Dispase Ⅱ at 270 rpm at 37 ℃ for 1 hour.Then homogenize tissue with a syringe (19G needle) and the resulting homogenous cell suspensions were filtered through a 70 μm cell strainer.Spin down at 350g for 5 minutes. Remove s...
Determination of total flavonoid content
Q&A Published: Aug 17, 2026Determination of Total Flavonoid Content in Choy Sum (Brassica chinensis var. parachinensis) Leaf Tissue Using a Microplate ReaderSubramanian Deivanai1 and Srinivasan Ramachandran21. AGRI Centre, Republic Polytechnic, 9 Woodlands Ave 9, 738964 Singapore2. Temasek Life Sciences Laboratory Limited, Research Link, National University Singapore, Buona Vista, 117604 Singapore.*For correspondence: deivanai@rp.edu.sg AbstractEstimating total flavonoid content (TFC) is important because flavonoids are a...
Antibody binding and deposition of complement components on bacterial surface
Q&A Published: Aug 11, 2026FITC labeling of S.pneumoniae Protocol Aim: Labeling of bacteria with FITC Requirements Fluorescein isothiocyanate (FITC, isomer I; Sigma Chemicals Co)DMSO PBS1x or Bicarbonate buffer (0.1M NaHCO3) pH ~8.5; 84mg in 10ml MQ Assay Prepare 10 mg/ml FITC solution in DMSO (10mg in 1ml DMSO)Grow bacteria in 10 ml THY (~0.6 OD550) Transfer to 15 ml falcon tube and harvest bacteria by centrifugation at 3500 rpm, 10 min.Wash bacteria once with 5 ml bicarbonate b...
Anti-GFP nanobody Alexa594 conjugation
Q&A Published: Aug 10, 2026Materials1) TCEP HCl, Cat#75259, Sigma-Aldrich2) P6 spin-column (Cat# 7326227, BioRad)3) Phosphate Buffered Saline without calcium chloride and magnesium chloride, Cat#D8537, Sigma-Aldrich)4) Alexa 594 maleimide (Cat#A10256, Invitrogen)5) Superdex 200 ProceedureStart with an eppendorf tube containing a solution of nanobody (100 micrograms/0.1 mL), add 1.5 microL of tris-(2-carboxyethyl)phosphine (1 M TCEP HCl, Cat#75259, Sigma-Aldrich). The final concentration of TCEP is 15 mM.Shake the mixture...