PLNT 2530 Lab -  Planning2023

updated Jan. 24, 2023


Scheduled lab sessions

Week

Agro I
Regeneration
Genomic DNA
Agro II
Dec. 5
make media for Agro
get Agro cells from Sonia
streak out and isolate colonies on Kanamycin, and non-Kan control plates



Dec. 12

inoculate liquid cultures from colonies
split into aliquots
alkaline plasmid prep of pI121 from Agro



Dec. 19

plant tobacco
RE digests of pI121 to confirm identity of plasmid



Dec. 25

Disha leaves









Jan. 9

make MS plates for transformation
sterilize materials



Jan. 16

Disha returns





Transform A.t. with pBI121; verify KanR
do plasmid prep to verify DNA
make glycerol stocks for -80
Prepare materials and media for Agro transformation



Feb. 6,7

 inoculate leaf discs with Agro






prepare media and supplies


Feb. 13,14


Plant Regeneration Lab session
plant transgenic tobacco and non-transgenic controls







Feb. 27



TAs run through
CTAB DNA extraction
nanopore quantification
RE digest, verify on gel
PCR, verify on gel







March 6




TAs run through
GUS assay
March 13



CTAB DNA extraction
nanodrop quantification of DNA

March 20



RE digest
start PCR
Run gel of RE DNA

March 27



run gel of PCR products






April 3

final photos of transformants


GUS assay of leaves
April 11


final photos of regenerating plants

 

April 12


report due



April 25



report due
report due