Multi-channel pipettes that can fit 20uL (0.5-10uL LTS) , 200uL (20-200uL LTS or Universal) and 1000uL (100-1200uL LTS or Universal) filter tips
20 uL tips: At least 8 boxes per plate (1 box per round of IP for adding serum to peptidome/lysate, 1 box for diluting phage lysate, 2 boxes for transferring lysate into PCR I and PCR I into PCR II, 1 box for PCR II barcoding primers, and 3 rows of 12 for adding PCR I MM into plate, PCR II MM into plate, and pooling barcoded PCR II products)
200 uL tips: At least 4 boxes per plate per round of IP (1 for adding AG beads to peptidome+serum, 1 for transferring beads in LB to E. coli, 1 for transferring lysate into 96-well PCR plate for library prep, and 3 rows of 12 for adding LB to beads, adding NaCl to lysate, and adding water to plate to dilute phage lysate for library prep)
1000 uL tips: At least 7 boxes per plate per round of IP (6 for RIPA washes, 1 for transferring lysate to new blocked plate, and 1 row of 12 for aliquoting BSA, 1 row for adding peptidome, 5 rows for adding RIPA for washes, 1 row for aliquoting E. coli)
Pipets (Universal or LTS is fine)
Rocker in 4°C walk-in fridge
37°C shaker(s) that can accommodate 125mL and 1L flasks and 2mL deep well plates
Magnetic rack(s) that can fit 15mL, 50mL conicals and 1.5mL tubes
Spectrophotometer that can measure OD600 of E.coli w/ cuvettes
96-Well Microtiter Plate Magnetic Separation Rack (NEB Cat# S1511S)
MicroAmp Adhesive Film Applicator (Thermo Scientific Cat# 4333183)
Foil seals (Bio-Rad Cat# MSF1001)
Rubber 96-well Imperma Mats (Genesee Cat#22-552)
0.22um PVDF filter 1L Stericups (Millipore Cat#S2GPU11RE)
Round 96 well, 2mL plates (Genesse Science Cat# 22-485)
Square 96 well, 2mL pyramid bottom plates (VWR Cat# 75870-796)
Air-O-Seal gas permeable seals (Thomas Scientific Cat#1149R80)
Hard-Shell 96-Well Polypropylene PCR Plates, Low Profile, Thin Wall, Skirted (Bio-Rad Cat# HSP9601)
20uL, 200uL and 1000uL LTS filter tips
Invitrogen Qubit Assay Tubes (Invitrogen Cat# Q32856)
Pierce 20X TBS Tween 20 Buffer 500mL (Thermo Scientific Cat# 28360)
Bovine Serum Albumin Fraction V (Sigma-Aldrich Cat# 10735094001)
AccuGENE 1 M Tris HCl Buffer (Lonza Cat# 51237)
TERGITOL solution Type NP-40 (Sigma-Aldrich Cat# NP40S-100ML)
SDS Solution, Molecular Biology Grade (10% w/v) (Promega Cat# V6553)
Triton X-100 (Sigma-Aldrich Cat# X100-100ML)
Glycerol for molecular biology, >99% (Sigma-Aldrich Cat# G5516-100mL)
1M HEPES (Thermo Scientific Cat# 15630080)
NaN3 (Sigma-Aldrich Cat#S2002-100G)
10X PBS -Ca and -Mg (Thermo Scientific Cat# AM9624)
Corning 1 L Millers LB Broth Corning Cat# 46-050-CM
BLT 5403 E. coli glycerol stock (EMD Millipore Cat# 69142-0.2ML; stored in -80; upon first use, make multiple personal glycerol stocks from first culture you make - don't keep taking from the lab stock)
Titered phage library (Appropriate titer for human peptidome library is at least 1 x 1010 pfu/mL. Titer decreases by ~1 log every 3 months that a library is stored at 4°C. A stored library should be re-titered before use if stored for more than a few months.)
Glial Fibrillary Acidic Protein, Polyclonal antibody (Agilent Cat# Z033429-2)
DynaBeads Protein A for Immunoprecipitation (Thermo Scientific Cat# 10008D)
DynaBeads Protein G for Immunoprecipitation (Thermo Scientific Cat# 10009D)
Nuclease-Free Water (Thermo Scientific Cat# AM9937)
Phusion High-Fidelity DNA Polymerase (NEB Cat# M0530L)
Deoxynucleotide (dNTP) Solution Mix (NEB Cat# N0447L)
Flag and Strep pooled primers
TruSeq Dual Index Sequencing Primers in 96-well plates
AMPure XP beads (Beckman Coulter Cat# A63881)
200 Proof Ethanol (Koptec Cat#64-17-5)
Qubit 1X dsDNA HS Assay Kit (Thermo Scientific Cat# Q33230)
Agilent High Sensitivity DNA Kit (Agilent Cat# 5067-4626)