谁帮我把这段文章翻译成英文，翻译好的多给分！！！！！！！！！！！！！！

Question

采用Sanger双脱氧链末端终止法，按T7 Sequencing TM Kit使用说明操作。[n一 P]
dATP标记。8% 变性聚丙烯酰胺凝胶，厚度0．4mm，胶长55cm，电泳在Pharmach LKB公司的Macrophor测序仪上进行，1200V~1500V，2～5小时。测序产物置x
光负片爆光，于一20℃过夜，放射自显影。
AT PJtts基固克隆与表达 将pPGAP —AT用NcoI—SalI双酶切，插入pBV220中，得到pBV220一a1一AT重组质粒。再将其中的a1-AT(BamHl—Ava1)片段用PCR定点诱变后的突变片段(BamHI—AvaI)置换，即得到垒基目的pBV220 AT P[tts表达质粒。该
质粒转化大肠杆菌HB101，在LB培养基中30"C生长至对数中期．42"C诱导3小时。收集菌体，超声破碎后分别用1mol／L，2mol／L尿素洗涤，8mol/L尿素溶解
包涵体。

Answer 1

比较靠谱的翻译：
采用Sanger双脱氧链末端终止法，按T7 Sequencing TM Kit使用说明操作。[n一 P] dATP标记。8% 变性聚丙烯酰胺凝胶，厚度0．4mm，胶长55cm，电泳在Pharmach LKB公司的Macrophor测序仪上进行，1200V~1500V，2～5小时。测序产物置x光负片爆光，于一20℃过夜，放射自显影。
Sanger’s dideoxynucleotide chain terminating method was adopted and conducted according to the application instructions of T7 Sequencing TM Kit. Marked as [n一 P] dATP. The electrophoresis of 8%denaturing polyacrylamide gel with 0.4mm thickness and 55cm in length was conducted on Pharmach LKB Company’s Macrophor DNA sequencing machine at 1200V~1500V for 2～5 hours. The sequenced product was then subject to X-ray negative film explosion light and autoradiography for overnight at - 20°C.

AT PJtts基固克隆与表达: 将pPGAP —AT用NcoI—SalI双酶切，插入pBV220中，得到pBV220一a1一AT重组质粒。再将其中的a1-AT(BamHl—Ava1)片段用PCR定点诱变后的突变片段(BamHI—AvaI)置换，即得到垒基目的pBV220 AT P[tts表达质粒。
AT PJtts gene cloning and expression: NcoI—SalI double enzyme digestion was conducted on pPGAP —AT, and it was inserted into pBV220 vector to obtain pBV220一a1一AT recombinant plasmid. One of the a1-AT(BamHl—Ava1) gene fragments was displaced by (BamHI—Ava1) which was a mutated fragment from PCR site directed mutagenesis process, and thus barrier based purpose pBV220 AT P[tts expression plasmids were obtained.

该质粒转化大肠杆菌HB101，在LB培养基中30"C生长至对数中期．42"C诱导3小时。收集菌体，超声破碎后分别用1mol／L，2mol／L尿素洗涤，8mol/L尿素溶解
包涵体。
The plasmids were transformed into Escherichia coli HB101, they were cultured in LB medium at 30"C until mid-logarithmic growth and induced for 3 hours at 42"C. The thalli were collected and subject to ultrasonication; they were separately cleansed by using 1mol／L and 2mol／L urea, and then dissolved in 8 mol/L urea.

【英语牛人团】

Answer 2

Using Sanger dideoxy chain termination method using T7 Sequencing TM Kit according to the instructions. [N a P]
dATP labeled. 8% polyacrylamide gel, the thickness of 0.4mm, plastic long 55cm, electrophoresis in Pharmach LKB's Macrophor sequencer on, 1200V ~ 1500V, 2 ~ 5 hours. Sequencing product set x
Negative light exposing, in a 20 ℃ overnight autoradiography.
AT PJtts Cloning and expression of the solid base pPGAP-AT with the NcoI-SalI double digestion, insert pBV220, the AT received pBV220 a a1 a plasmid. Then one of a1-AT (BamHl-Ava1) after site-directed mutagenesis with PCR fragments of mutated fragments (BamHI-AvaI) replacement, that is to be barrier-based purpose pBV220 AT P [tts expression plasmid. That
Plasmid was transformed into E. coli HB101, in LB medium 30 "C to the logarithmic growth of the mid-.42" C-induced 3 hours. Collection of biomass, respectively, after sonication with 1mol / L, 2mol / L urea washing, 8mol / L urea solution
Inclusion bodies.

Answer 3

采用Sanger双脱氧链末端终止法，按T7 Sequencing TM Kit使用说明操作。[n一 P]
Using Sanger dideoxy chain termination method, according to the T7 Sequencing TM Kit usage instructions. [ n P]
dATP标记。8% 变性聚丙烯酰胺凝胶，厚度0．4mm，胶长55cm，电泳在Pharmach LKB公司的Macrophor测序仪上进行，1200V~1500V，2～5小时。测序产物置x
DATP markers. 8% denaturing polyacrylamide gel, gel thickness 0.4mm, long 55cm, electrophoresis in Pharmach LKB's Macrophor sequencing instrument, 1200V ~ 1500V, 2 ~ 5 hours. The sequencing of the X product
光负片爆光，于一20℃过夜，放射自显影。
Light negative exposure, in a 20 degrees overnight, autoradiography.
AT PJtts基固克隆与表达 将pPGAP —AT用NcoI—SalI双酶切，插入pBV220中，得到pBV220一a1一AT重组质粒。再将其中的a1-AT(BamHl—Ava1)片段用PCR定点诱变后的突变片段(BamHI—AvaI)置换，即得到垒基目的pBV220 AT P[tts表达质粒。该
AT PJtts based solid cloning and expression of pPGAP - AT NcoI - SalI double digestion, insert pBV220, pBV220 A1 AT recombinant plasmid. And then one of the a1-AT ( BamHl - Ava1 ) fragment with PCR site-directed mutagenesis of the mutant fragment ( BamHI - AvaI ) replacement, namely base base to pBV220 AT P [ TTS expression plasmid. The
质粒转化大肠杆菌HB101，在LB培养基中30"C生长至对数中期．42"C诱导3小时。收集菌体，超声破碎后分别用1mol／L，2mol／L尿素洗涤，8mol/L尿素溶解
Plasmid was transformed into E. coli HB101, in LB medium 30" C growth to the logarithmic term . 42" C induced by 3 hours. Lactobacillus collection, ultrasonic crushing respectively after using 1mol / L, 2mol / L urea washing, 8mol / L dissolving urea
包涵体。
Inclusion body.

Answer 4

The Sanger double deoxidizing chain end, according to terminate method T7 Sequencing TM Kit instructions operation. [n a P]
DATP mark. 8% degeneration polyacrylamide gel, thickness of 0.4 mm, 55 cm long, electrophoresis glue in Pharmach LKB company Macrophor sequencing instrument, 1200 V ~ 1500 V, 2 ~ 5 hours. Sequencing product for x
The light of the negative light, in a critical 20 ℃ for the night, radiation from enhancement.
AT PJtts base solid cloning and expression will pPGAP-AT NcoI-SalI with double enzyme, insert pBV220 cut, get pBV220 a a1 AT a restructuring plasmid. Then one of the a1-AT (BamHl-Ava1) clips with the mutation PCR site-directed mutagenesis footage (BamHI-AvaI), that is, get base and replacement pBV220 purpose AT P [TTS expression plasmid. This
Plasmid into e. coli HB101, the medium in langmuir-blodgett 30 "C growth to mid logarithm. 42" C induction of three hours. Collect bacteria, ultrasonic broken respectively with 1 mol/after L, 2 mol/L urea washing, 8 mol/L urea dissolve
Inclusion body

Answer 5

翻译

The Sanger double deoxidizing chain end, according to terminate method T7 Sequencing TM Kit instructions operation. [n a P]
DATP mark. 8% degeneration polyacrylamide gel, thickness of 0.4 mm, 55 cm long, electrophoresis glue in Pharmach LKB company Macrophor sequencing instrument, 1200 V ~ 1500 V, 2 ~ 5 hours. Sequencing product for x
The light of the negative light, in a critical 20 ℃ for the night, radiation from enhancement.
AT PJtts base solid cloning and expression will pPGAP-AT NcoI-SalI with double enzyme, insert pBV220 cut, get pBV220 a a1 AT a restructuring plasmid. Then one of the a1-AT (BamHl-Ava1) clips with the mutation PCR site-directed mutagenesis footage (BamHI-AvaI), that is, get base and replacement pBV220 purpose AT P [TTS expression plasmid. This
Plasmid into e. coli HB101, the medium in langmuir-blodgett 30 "C growth to mid logarithm. 42" C induction of three hours. Collect bacteria, ultrasonic broken respectively with 1 mol/after L, 2 mol/L urea washing, 8 mol/L urea dissolve
Inclusion body.

Answer 6

Using Sanger dideoxy chain termination method, according to the T7 Sequencing TM Kit usage instructions. [ n P]DATP markers. 8% denaturing polyacrylamide gel, gel thickness 0.4mm, long 55cm, electrophoresis in Pharmach LKB's Macrophor sequencing instrument, 1200V ~ 1500V, 2 ~ 5 hours. The sequencing of the X productLight negative exposure, in a 20 degrees overnight, autoradiography.AT PJtts based solid cloning and expression of pPGAP - AT NcoI - SalI double digestion, insert pBV220, pBV220 A1 AT recombinant plasmid. And then one of the a1-AT ( BamHl - Ava1 ) fragment with PCR site-directed mutagenesis of the mutant fragment ( BamHI - AvaI ) replacement, namely base base to pBV220 AT P [ TTS expression plasmid. ThePlasmid was transformed into E. coli HB101, in LB medium 30" C growth to the logarithmic term . 42" C induced by 3 hours. Lactobacillus collection, ultrasonic crushing respectively after using 1mol / L, 2mol / L urea washing, 8mol / L dissolving ureaInclusion body.