Thermo Scientific OWL HEP-1, Руководство по эксплуатации и техническому обслуживанию

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3-2 Semidry Electroblotter Thermo Scientific
Section 3
Using the System
Transfer Settings
(continued)
Read your power supply's instructions to ensure that the power supply will
work at a voltage lower than lOV. These voltages often occur in semidry
blotting. Contact the manufacturer regarding the unit's performance under
high current, low voltage conditions if you have any questions.
While general conditions can be described which will result in successful
transfer of most molecules, it should be noted that optimal transfer
conditions will vary based on the characteristics of the molecule you are
working with. Some factors that affect transfer rate and efficiency include
molecule size, charge, gel thickness and percentage, and hydrophobicity.
The reference list at the end of this manual provides useful information
that can help you choose optimal conditions for efficient transfer of a
specific molecule.
Factors That Affect
Transfer Efficiency
Protein
HEP-3/ Sequencing
gel - see HEP-1 set-
tings for other size
gels on this unit
DNA/RNA
Filter Paper
FP-1, 20 x 20cm, FP-6,
9 x 9cm, FP-4, 10 x
10cm, FP-7, 12 x 16cm,
or FP-5, 18 x 20cm FP-2, 35 x 45cm, or
FP-3, 46 x 57cm
FP-1, FP-4, FP-5, FP-6, or
FP-7
Membrane
Nitrocellulose .45 or
0.2μ, PVDF 0.45μ or
0.2μ Nylon Nylon
Transfer Buffer
Towbin Buffer, Buffer
Electroblot Buffer Kit,
ER-35, 3 Buffer System
Bjerrum and Schafer-
Nielsen
0.5X-1X TBE 0.5X-1XTBE, TAE, NAQ
Power Supply OSP-135 or equivalent
High current/low volt-
age power supply
OSP-135 or equivalent
Power Settings
Constant current 0.8-
3mA per cm2 gel sur-
face area 10-14 Volts
maximum
For entire gel, 1200-
1400 mA approximate-
ly 0.8mA/cm2/
produces ~5-8V
Constant Current 0.5-
3mA/cm2 gel surface
area 10-14 Volts maxi-
mum
Running Time
30 minutes to 2 hours
Needs to be experi-
mentally determined
(large molecules need
longer transfer times)
15-20 minutes at this
setting
30 minutes to 2 hours,
generally in the lower
range
Running Conditions