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Methods

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AGROBACTERIUM TRANSFORMATION

Agrobacterium Transformation: Freeze-Thaw (posted 2/13/18)

Agrotransformation

Conjugation of Agro with Helper Strain

Silwet/Synperonic

BACTERIAL TRANSFORMATION

Heat Shock Transformation of E. coli competent cells (prepared by Tamara Miller, 5/27/10)

E. coli Competent Cell Preparation (prepared by Han Qi Tan, 7/09)

Bacterial Transformation (MJC)

Bacterial Culture Preparation/Bombardment (liquid culture)

Bacterial Media
 YEP medium
 Agro LB medium
 LB medium

Competent Cells (MJC)

BOMBARDMENT

Preparation of Gold

Procedure for Helium Bombardment

Helium Bombardment (MJC)

Particle Prep (MJC)

SORGHUM TRANSFORMATION

Clemente Protocol (posted 4/19/10)

TX430 Sorghum Transformation Protocol (posted 9/21/12)

Selection of heads for IEs (posted 6/7/10)

Regeneration

Storage and Handling of G418

SETARIA TRANSFORMATION

Agrobacterium tumefaciens-mediated Transformation of Setaria viridis (posted 9/21/12)

CONVERTING SCOPE IMAGES TO JPEGs

DNA ISOLATION

Genomic

Miniprep - Urea Extraction Method (prepared by Ling Meng, December 2012).

Plant Chloroplast Isolation - with tobacco specific modifications (prepared by Elyssa Lewis, November 2012)

CTAB DNA Isolation from Amit Dhingra, September 2012)

Damon’s All Natural (no organics) Genomic Miniprep (prepared by Damon Lisch, June 2009)

Hybrid (Natural & Phenol) DNA Extraction for Sorghum

Plasmid
 Procedure for sending binary plasmid DNA via standard mail

10 Min Miniprep

10% SDS Solution (posted 3/16/10)

TENS – Solution used for plasmid or cosmid isolation from bacteria (via 10 minute miniprep protocol) (posted 3/16/10)

MJC protocols
 10 Min Miniprep (MJC)

Genomic DNA Extraction (MJC)

Maxi Prep (MJC)

Seed DNA Extraction (MJC)

Urea Miniprep (MJC)

DNA MANIPULATION

DNA Ligation (MJC)

Ligation Transformation (MJC)

In situ protocol (MJC)

PCR Purification (MJC)

Composition of restriction enzyme reaction buffers (posted 4/20/10)

Extended digests with restriction endonucleases (posted 4/20/10)

DNA PURIFICATION & QUANTIFICATION

DNA Quantification (MJC)

Gel Extraction (MJC)

QIAEX II Agarose Gel Extraction (MJC)

Spin Miniprep (MJC)

Wizard DNA Purification (MJC)

GEL ELECTROPHORESIS

TAE, TBE Recipes

GFP SCREEN

Screening T3 seeds (Prepared by Karan Verma, June 2009)

Instructions for Xenogen machine (posted 3/14/11)

GRINDING

Protocol for using Wiley Mill (posted 1/26/18)

GUS SCREEN

GUS: Histochemical Staining with X-Gluc (posted 3/9/10)

Membrane Hybridization Protocol

Storage of X-Gluc

Rueb, S. and Hensgens, L.A.M. 1989. Improved histochemical staining for beta-D-glucuronidase activity in monocotyledonous plants. Rice Genetics Newsletter, Vol. 6, December 1989.

IMMATURE EMBRYO ISOLATION

Immature Embryo Isolation - Wheat or Sorghum

Immature Embryo Isolation (MJC)

MARKER DETECTION

Whole plant NPTII assay

MICROSPORES

Root tip squash protocol

PCR

PCR Polymerase Chain Reaction Protocol for PMI gene

Printing/saving pictures from the GelDoc

PROTEIN

Western blot protocol (prepared by Emily Kao, posted 2/6/12).

STARCH

EnzyChrom Starch Assay Kit (posted 2/1/18)

RNA PROTOCOLS

mRNA Extraction from Callus Protocol (Adapted by Tom Kleist from Chang, Puryear, & Cairney 1993. Posted 3/8/10)

cDNA Synthesis Protocol (posted 3/8/10)

PLANT & PLANT CARE

Hoagland's solution

Planting

STANDARD OPERATING PROCEDURES (SOPs)

Autoclaves
 Ethidium Bromide
 Gloves
 Phenol
 SDS-Polyacrylamide Gel Electrophoresis

SOUTHERN BLOT

Nonradioactive DIG Southern (prepared by Tamara Miller, posted 9/20/12).

Southern Blot - Making & Running Gels

Southern Blot – Membrane Transfer

Nonradioactive Southern Blot Protocol (prepared by Tamara Miler, posted 8/3/12)

Product info:
anti-DIG-AP
 CDP-Star ready to use
 DIG Easy hyb granules
 PCR DIG labeling mix
 Roche N pos membrane