- High
throughput transformation without making competent cells
- Simultaneous
transformation of recovered library vectors and bait when processing
putative positive two-hybrid clones.
|
|
The EZ-Yeast
Transformation kit is designed for high throughput transformation with
an efficiency of up to 1000 colonies per µg plasmid DNA. For high
efficiency transformation use the Alkali Cation Yeast Transformation Kit or the Yeast
Spheroplast Transformation kit.
Shipping
& Storage
Ambient
Transformation
efficiency at different temperatures
| Vector(s) |
overnight
Room temp |
30
min
30°C |
30
min
37°C |
30
min
42°C |
|
| pVA3 (bait) |
38 |
40 |
100 |
300 |
| pTD1 (library) |
4 |
19 |
25 |
100 |
| bait library |
0 |
2 |
8 |
21 |
The yeast
two-hybrid reporter strain SFY526 is transformed as per protocol with
incubation at the indicated conditions. The number of transformants after
3 days at 30°C is shown. Bait, library and bait library transformants
are plated on SDA-TRP (cat.# 4812-), SDA-LEU-TRP (cat.# 4811-) and SD-LEU-TRP
(cat.# 4823-) respectively.
 Simultaneous
transformation of recovered library and bait vectors
| Yeast
two-hybrid reporter strains |
| Vectors |
SFY526 |
HF7C |
Y190 |
EGY48 |
|
| Bait library |
18 |
34 |
16 |
13 |
The various
yeast two-hybrid reporter strains are transformed simultaneously with
bait and library vectors as per protocol (30 min at 42°C). The number
of transformats after 3 days at 30°C is shown. Bait and library vectors
for SFY526, HF7C and Y190 are pVA3 and pTD1; cotransformants are plated
on SD-TRP-LEU (cat.# 4823-). Bait and library for EGY48 are pEG202-P53
and pJG4-5-LTA; cotransformants are plated on SD-HIS-TRP (cat.# 4826-).
The
Protocol, in brief:
- Transfer
yeast colonies or cells form overnight culture to sterile 1.5ml tubes
or 96 well plates.
- Add
vector, carrier DNA, EZ-Transformation Solution and mix.
- Incubate
30 min at 42°C.
- Plate
the cells on selective media.
- Transformants
are observed after 2-3 days.
| Cat
No. |
Size |
|
| 2100-200 |
200 preps |
|
