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Table 1 Effects of plant material on protoplast isolation on protoplast transfection.

From: Tape-Arabidopsis Sandwich - a simpler Arabidopsis protoplast isolation method

Light condition

Method

Protoplast No. (×107)

Transformation efficiency (%)*

Broken (%)

Low

Cutting

1.8 ± 0.3

64.8 ± 5.8

1.7 ± 2.9

Low

Tape

1.4 ± 0.2

66.2 ± 8.4

3.2 ± 5.6

Optimal

Cutting

3.7 ± 1.3

53.0 ± 4.0

24.9 ± 10.3

Optimal

Tape

3.0 ± 0.3

60.7 ± 3.2

1.8 ± 3.3

  1. One gram of leaves derived from plants was grown under normal (optimal) or low-light conditions [4] were used. Protoplasts were isolated and purified using Tape-Arabidopsis Sandwich and cutting protocols. Total cell number (intact and broken cells) was calculated. The 'Broken (%)' is: 100 × (broken cell number/total cell number). The broken protoplasts were not counted during the transfection efficiency calculation. The transfection efficiency is the percentage of intact protoplasts with fluorescent/intact protoplasts. These protoplasts were transformed with VirE2-Cerullean. Each treatment was replicated three times (n = 3).
  2. *: % of intact protoplasts showing VirE2-Cerulean fluorescence.