Protocols

Most of these protocols are from other labs or from online sources. Some were modified by the Franks' lab for our specific purposes. Questions regarding any of these protocols please contact us at: rgfranks@ncsu.edu





Agarose gels

  1. Agarose gel
  2. MetaPhor gel

Antibody purification

  1. From Zhongchi’s Lab

Artificial miRNAs

  1. Cloning of artificial microRNAs (Schwab)
  2. Schwab Weigel synmiRNA

CAPS_SSLP Markers

  1. ANT alleles
    1. ant-1 caps
    2. ANT-3 SSLP
    3. ANT-8 CAPS
    4. ANT__9
  2. ATM 49 Mapping
    1. K14B20-Nde I
    2. F6H11-Alu I
    3. K14B20-NdeI 65890
    4. K14B29-Spe I
    5. MPA24-Ase I
  3. CRC
    1. Crc-1 dCAPS
    2. Crc-1 Mutant sequence
  4. Katanin p80 salk lines
    1. Salk_041812
    2. Salk_042104
    3. Salk_078009
    4. Salk_123263
  5. PHB and REV Alleles
    1. Phb-1D
    2. Rev5
  6. SEUSS alleles
    1. Seu-1 CAPS
    2. Seu-2 CAPS
    3. Seu-3 CAPS
    4. Seu-5 CAPS
  7. SLK alleles
    1. Sequence of SLK1_6A
    2. SLK1_6A
    3. SLK1_11D
    4. SLK1_043166
    5. SLK1_801304
    6. SLK1_852593
    7. SLK1_093829C
    8. SLK2_CS842569
    9. SLK2_038662
    10. SLK2_085761C
    11. SLK2_089954
    12. SLK3_090714
    13. SLK3_117317
    14. SLK3_CS811046
  8. SUM alleles
    1. Sum45-1 CAPS
    2. SUM132 dCAPS

CEL 1 assay

  1. 4597

ChIP

  1. ChIP protocol magnetic beads
  2. ChIP protocol with 2ndary antibody

Chloral Hydrate Clearing

  1. Chloral Hydrate

DAPI GFP labeling

  1. DAPI labeling of plant roof hair
  2. GFP FIX protocol
  3. GFP fix with Proidium Iodide
  4. Haseloff99b
  5. Staining arabidopsis root

DNA preps

  1. Alonso Lab 96-well Plant DNA extraction
  2. Franks Lab 96-well DNA
  3. Leaf preps Edwards (few samples)
  4. Phb1d small quantity leaf preps
  5. Single Embryo PCR

FTA cards

  1. BC01 Purification for AFLA analysis
  2. BD05 Applying on FTA cards
  3. BD08 Preparing Disc for Analysis
  4. BD09 Punching protocol
  5. BD10 Purificiation RFLP analysis

GUS Staining

  1. Franks lab GUS protocol

Histochemistry

  1. DAPI staining
  2. GFP fix for confocal (Franks lab)
  3. Toluidine Blue Staining
  4. Johansen’s Saffranin
  5. Slide Stain Guide

Image Analysis/Publication Figures

  1. Making figures for publication

Immuno Histo Chemistry

  1. antibody protocol
  2. Immunoab purification
    1. Immuno histochemistr fixation
    2. Robertson Vibratome imunohisto
    3. Uli M proto_immuno

InSitu

  1. Institu probes
  2. Institu protocol
  3. PVA insitu
  4. PVA solutions

Media

  1. 1/2 MS medium
  2. LB medium
  3. Surface sterilization of Arabidopsis seeds

Microscopy

  1. Agarose Micrtome for L CM
  2. Compound scope (DIC and Floresence)

Nanodrop

  1. Nanodrop spectrophotometer

Oligo design

  1. Salk conform oligos

Propidium Iodide

  1. Mark Running PI confocal protoocol

Protein Expression

  1. Recombinat protein expression test

qRT PCR

  1. plate grid
  2. qRT CR-ABI 7900 instructions

RNA preps

    1. Qiagen
    2. dissected carpels
    3. Trizol

Selection of transformants

  1. hygromycin selection

SEM

  1. materials for SEM
  2. Protocol from Valerie
    1. Fang Bao
    2. Fumehood
    3. Working Safely GTA

Sequencing

  1. Enzymatic Cleanup of PCR
  2. sequencing from amplicon

Stocks and Strains

  1. Bacterial strains
  2. Constructs and strains form

TAQ enzyme purification

  1. 10x Taq Buffer
  2. Taq Purification -Alonso Lab

Transformation

  1. Arabidopsis floral dip PDF
  2. Arabidopsis floral dip transformation
  3. Clough and Bent floral dip PDF
  4. Electro-Competent Cells
  5. Electroporation
  6. Heat Shock transformation
  7. Large DNA transformation PDF
  8. Simple Arabidopsis dip

Vector maps

  1. Cyclin promoter map
  2. PBI101.3 TDNA Map
  3. PCRII topo map

Western blotting

  1. Dot Blot protocol
  2. Protein isolation
  3. SDS Page gel (franks)
  4. SDS PAGE gel - Bowtell
  5. Western blot
  6. Western blot using AP

Yeast-2 Hybrid

  1. Y2H reference Gietz MCB v172_PDF
  2. Filter lifting assay
  3. Long term storage of stocks
  4. ONPG assay
  5. Transformation

Other

  1. Save a snowflake for Decade