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Total RNA was isolated using TBlzol reagent (Invitrogen ! fe
Technologies, Carlsbad, CA, USA) according to the mar facturer's
instructions. The purity and concentration ofthe BRNAw /e
Jme
spectrophotometer (Maestrogen, Las Vegas, NV, USA), and cDNAs
Were synthesized using the miScript II RT kit (Qiagen,
Germany) according to the manufacturer's instructions.
evaluate the expression of MMP-1, quantitative PCR was
performed using the following primers: forward, 5-TCT
GIAGGTTGATCCCAGAGAGCAG-3' and reverse, 5-CAGGG
TGACACCAGTGACTGCAC-3“ using EvaGreen dye (Solis
BioDy Line-Gene K software (Bioer
Technology Co,, Ltd., Hangzhou, China). The Ct value for each
gene was normalized to B-actin using the following primers:
forward, 5“-GGATTCCTATGTGG 3 and reverse, 5/-
GGCTCGGGTGAGGAITCTTCATG-3/ ive expression levels
Of each gene were calculated using the 2-AAct method, as
previously described-
RNA萃取&定量PCR
根據製造商的說明,使用
分離總RNA。使用Ma ) Maestrog
對RNA的純度和渡度進行了評價,根據製造商的說明。使用iS
(Qiagen) 合成了cDNAs。為了評估MMP-1的表達。使
用以下引物(子)進行了定量PCR :
正向5-TCTGACGTTGATCCCAGAGAGCAG-3'和
向5-CAGGGTGACACCAGTGACTGCAC-3', 使用EvaGreen染料
ss BioDyne) 和Line-GeneK軟件。使用以下引物將每個基因的@
B-actin
向$:-GGATTCCTATGTGGGCGACGA-3',
CGCTCGGTGAGGATCTTCATG-3'。 如前所述,使用2-se'法
LuyittogenT 生 人 oo
