Geogosin-based randomly labeled DNA probes.
-
Last Update: 2020-10-30
-
Source: Internet
-
Author: User
Search more information of high quality chemicals, good prices and reliable suppliers, visit
www.echemi.com
1. Mark
DNA
Probe Each standard reaction can mark 10ng to 3linear DNA, or a larger amount of DNA, but all ingredients and volumes increase accordingly.(1) DNA probe hot denaturation, boiling 10min, quickly cooled in ice/ethanol more than 5min, to be used.
(2) take Eppendorf tube (1.5 ml) and place it on ice, add the following and
reagents
:
freshly denatured DNA 1-3
he6-poly
nucleotides
mixture 2 (tube 5)
d NTP marker with mixed substrate 2 (tube 6)
plus sterile heavy steamed water to 19
DNA polymerase I. large fragment (Klenow) 1 (tube 7)
(3) insulation at 37 degrees C at least 60min, up to 20h.
(4) boil 5min to terminate the reaction.
(5) 15000rpm centrifuge 30s and place on ice for use.
2. Pre-hybridization According to 100cm2 membrane with 20 to 40 ml pre-hybrid solution, pre-hybridization makes the solution in a flowing state.
prebreeding liquid composition: 5㗓SC
0.5% (W/V) sealing reagent (tube 11)
0.1% (W/V) N-12 Sodium alkyl creatinine
0.02% (W/V)
SDS
membrane into a plastic bag, filled with pre-hybrid liquid, removed the gas after sealing, pre-hybridized at 65 degrees C overnight.
.
This article is an English version of an article which is originally in the Chinese language on echemi.com and is provided for information purposes only.
This website makes no representation or warranty of any kind, either expressed or implied, as to the accuracy, completeness ownership or reliability of
the article or any translations thereof. If you have any concerns or complaints relating to the article, please send an email, providing a detailed
description of the concern or complaint, to
service@echemi.com. A staff member will contact you within 5 working days. Once verified, infringing content
will be removed immediately.