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Chen et al. Screening of genetic loci

Table 1: Primer sequences for scanning microsatellite loci on the 17th chromosome of mouse genome
Loci name Primer sequence Genetic distance (cM)
D17MIT245.1 Forward FAM-TGTGCTCTGGCTAGGGAGTT 3
Reverse CACATTCATATGTACACACACATGC
D17MIT143.2 Forward FAM-CTTACAAGCATCCTGTGGAACTC 5
Reverse GAGGACCAACAGTCAAACATAGC
D17MIT46 Forward FAM-TCCACCCCACTACCTGACTC 11.7
Reverse CCCTTCTGATGACCACAGGT
D17MIT146.1 Forward FAM-CTGTCAGCAGAACGTTCCTTAGT 17.1
Reverse CCAACTCAAGCCTTACATAGTGG
D17MIT51.1 Forward FAM-TCTGCCCTGTAACAGGAGCT 22.9
Reverse CTTCTGGAATCAGAGGATCCC
D17MIT10.1 Forward FAM-TGCACTTGCATAAGGAAAAC 24.5
Reverse GACTTTGGGGCCTACTTATG
D17MIT180.1 Forward FAM-AGACACTGTCTAAAAACACAAGATGG 29.4
Reverse TTGTGTTCATATGCATGTGTGC
D17MIT20.1 Forward FAM-AGAACAGGACACCGGACATC 34.3
Reverse TCATAAGTAGGCACACCAATGC
D17MIT184 Forward FAM-TGCACTACCCAAACATGCAT 38.5
Reverse ACTTCTGACAGGAAGCATCCA
D17MIT93.1 Forward FAM-TGTCCTTCGAGTGTTTGTGTG 44.5
Reverse TCCCCGGTGAATGAGTTATC
D17MIT39.1 Forward FAM-CCTCTGAGGAGTAACCAAGCC 45.3
Reverse CACAGAGTTCTACCTCCAACCC
D17MIT122.1 Forward FAM-TCTCTTCACTGCAATGGAACA 51.9
Reverse GAACCTATAGGCTCTTGAATAGATGG
homologous genes that showed some of the following Down syndrome critical region gene 1-like 1, MSM
characteristics: (1) containing many quantitative trait lymphoma resistance 1, etc.; (5) containing mouse
loci, such as epididymal fat pad weight quantitative tissue associated antigen H-2.
trait loci (QTL) 3, subcutaneous fat pad weight QTL 4,
spleen weight QTL 9, etc.; (2) containing some genes DISCUSSION
related to the important physiological functions of the
body such as Mut methylmalonyl-Coenzyme A mutase; It has been widely observed that different species or
(3) containing genes related to the developmental and even individuals of the same species show differences
physiological function such as early growth adjusted in response to infection, but the explanation for this
QTL 2, early growth QTL 5, etc.; (4) containing phenomenon is still rather controversial. There have
genes associated with some diseases such as the
Table 3: Microsatellite loci scanning using BALB/C and
Table 2: PCR reaction system and PCR reaction condition C57BL/6 inbred mice
PCR reaction system (total volume: 10 uL) Microsatellite loci BALB/C C57BL/6
Non-enzyme water 5.4 μL susceptible tolerant
10 × PCR buffer 1.0 uL D17MIT245.1 194 202
2+
Mg (25 mmol/L) 0.5 uL D17MIT143.2 112 112
dNTP (each 2.5 mmol/L) 1.0 μL D17MIT46 218 236
P1 (5 pM) 0.5 uL
P2 (5 pM) 0.5 uL D17MIT146.1 166 166
Template DNA (30-50 ng/uL) 1.0 uL D17MIT51.1 152 154
Ex-Taq enzyme (5 U/μL) 0.1 μL D17MIT10.1 155 155
PCR reaction condition D17MIT180.1 139 137
95 ℃ 5 min D17MIT20.1 163 175
94 ℃ 30 s
Time 30 s D17MIT184 126 128
72 ℃ 30 s D17MIT93.1 155 155
Repeat the 2nd to 4th steps for totally 38 cycles D17MIT39.1 86 104
72 ℃ 10 min
Store at 4 ℃ D17MIT122.1 141 141
PCR: polymerase chain reaction Microsatellite loci with difference were marked in bold
276 Neuroimmunology and Neuroinflammation ¦ Volume 3 ¦ December 26, 2016