六、常用凝胶的技术参数
1.葡聚糖凝胶的某些技术数据
种类 |
干颗粒直径(μ) |
分子量分级范围 |
床体积毫升/克干分子筛 |
得水值 |
溶胀最少平衡时间(h) | 柱头压力(kPa)(2.5cm直径柱) |
||
肽及球形蛋白质 | 葡聚糖(线性分子) | 室温 | 沸水浴 | |||||
Sephadex G-10 | 40~ 120 | ~700 | ~700 | 2~ 3 | 1.0±0.1 | 3 | 1 | |
Sephadex G-10 | 40~ 120 | ~1500 | 1500 | 2.5~3.5 | 1.5±3.5 | 3 | 1 | |
Sephadex G-25 | ||||||||
粗级 |
100~300
(≈5~100目) |
|||||||
中级 |
50~150
(≈100~200目) |
1,000~ 5,000 |
100~ 5,000 |
4~6 |
1.5±0.2 |
6 |
2 |
|
细级 |
20~800(≈200~400目) | |||||||
超细 |
10~40 | |||||||
Sephadex G-50 | ||||||||
粗级 |
100~200 | |||||||
中级 |
50~150 | 1,500~ | 500~ | |||||
细级 |
20~80 | 30,000 | 10,000 | 9~11 | 5.0±0.3 | 6 | 2 | |
超细 |
10~40 | |||||||
Sephadex G-75 | ||||||||
40~120 | 3,000~ | 1,000~ | ||||||
超细 |
10~40 | 70,000 | 950,000 | 12~15 | 7.5±0.5 |
24 |
3 |
3.92~15.86 |
Sephadex G-100 | ||||||||
40~120 | 4,000~ | 1,000~ | ||||||
超细 |
10~40 | 1,500,000 | 150,000 | 15~20 | 10.0±1.0 | 48 | 5 | 2.35~9.41 |
Sephadex G-150 | ||||||||
40~120 | 5,000~ | 1,000~ | 20~30 | |||||
超细 |
10~40 | 400,000 | 150,000 | 18~22 | 15.0±1.5 | 72 | 5 | 0.88~3.53 |
Sephadex G-200 | ||||||||
40~120 | 5000~ | 1000~ | 30~40 | |||||
10~40 | 800,000 | 200,000 | 20~25 | 20.0±2.0 | 72 | 5 | 0.39~1.57 |
2.聚丙烯酰胺凝胶的技术数据
型号 |
排阻的下限 (分子量) |
分级分离范围 (分子量) |
膨胀后的床体积(ml/g干凝胶) |
膨胀所需最少时间(室温,h) |
Bio-gel-P-2 | 1,600 | 200~2,000 |
3.8 |
2~4 |
Bio-gel-P-4 | 3,600 | 500~4,000 |
5.8 |
2~4 |
Bio-gel-P-6 | 4,600 | 1,000~5,000 |
8.8 |
2~4 |
Bio-gel-P-10 | 10,000 | 5,000~17,000 |
12.4 |
2~4 |
Bio-gel-P-30 | 30,000 | 20,000~50,000 |
14.9 |
10~12 |
Bio-gel-P-60 | 60,000 | 30,000~70,000 |
19.0 |
10~12 |
Bio-gel-P-100 | 100,000 | 40,000~100,000 |
19.0 |
24 |
Bio-gel-P-150 | 150,000 | 50,000~150,000 |
24.0 |
24 |
Bio-gel-P-200 | 200,000 | 80,000~200,000 |
34.0 |
48 |
Bio-gel-P-300 | 300,000 | 100~400,000 |
40.0 |
48 |
3.琼脂糖凝胶的技术数据
型号 |
琼脂糖含量 %(W/W) |
排阻的下限 (分子量) |
分级分离的范围(分子量) |
生产厂家 |
Sepharose 4B | 4 | 0.3×106~3×106 | Pharmacia | |
Sepharose 2B | 2 | 2×106~25×106 | ||
Sagavac 10 | 10 | 2.5×105 | 1×104~2.5×105 | Seravac |
Sagavac 8 | 8 | 7×105 | 2.5×104~7×105 | |
Sagavac 6 | 6 | 2×106 | 5×104~2×106 | |
Sagavac 4 | 4 | 15×106 | 2×105~15×106 | |
Sagavac 2 | 2 | 150×106 | 5×105~15×107 | |
Bio-gel A-0.5M | 10 | 0.5×106 | <1×104~0.5×106 | Bio-Rad |
Bio-gel A-1.5M | 8 | 1.5×106 | <1×104~1.5×106 | |
Bio-gel A-5M | 6 | 5×106 | 1×104~5×106 | |
Bio-gel A-15M | 4 | 15×106 | 4×104~15×106 | |
Bio-gel A-50M | 2 | 50×106 | 1×105~50×106 | |
Bio-gel A-150M | 1 | 150×106 | 1×106~150×106 |
4.各处凝胶所允许的最大操作压
凝胶 |
最大静水压(kPa) |
Sephadex |
|
G-10 |
9.8 |
G-15 |
9.8 |
G-25 |
9.8 |
G-50 |
9.8 |
G-75 |
4.9 |
5.琼脂糖凝胶浓度与线性DNA分辨范围
凝胶浓度(%) |
线性DNA长度(bp) |
0.5 |
1000~30000 |
0.7 |
800~12000 |
1.0 |
500~10000 |
1.2 |
400~7000 |
1.5 |
200~3000 |
2.0 |
50~2000 |
6.染料在变性聚丙烯酰胺凝胶中的迁移速度
凝胶浓度(%) |
溴酚蓝 |
二甲苯青FF |
5.0 |
35bp |
140bp |
6.0 |
26bp |
106bp |
8.0 |
19bp |
75bp |
10.0 |
12bp |
55bp |
20.0 |
8bp |
28bp |
7.染料在非变性聚丙烯酰胺凝胶中的迁移速度
凝胶浓度(%) |
溴酚蓝 |
二甲苯青FF |
3.5 |
100bp |
460bp |
5.0 |
65bp |
260bp |
8.0 |
45bp |
160bp |
12.0 |
20bp |
70bp |
15.0 |
15bp |
50bp |
20.0 |
12bp |
45bp |
七、氨基酸的特性
氨基酸名称 | 三字母缩写 | 单字母缩写 |
质量 |
侧链电离的pHa值 |
结构式
|
丙氨酸(alanine) | Ala | A | 89.09 | ||
精氨酸(arginine) | Arg | R | 174.2 | 12.48 | |
天冬酰氨(asparagine) | Asn | N | 132.1 | ||
天冬氨酸(aspartic acid) | Asp | D | 133.1 | 3.86 | |
半胱氨酸(systeine) | Cys | C | 121.12 | ||
谷氨酰胺(glutamine) | Gln | Q | 146.15 | ||
谷氨酸(glutamic acid) | Glu | E | 147.13 | 4.25 | |
甘氨酸(glycine) | Gly | G | 75.07 | ||
组氨酸(histidine) | His | H | 155.16 | 6.0 | |
异亮氨酸(isoleucine) | lle | I | 131.17 | ||
亮氨酸(leucine) | Leu | L | 131.17 | ||
赖氨酸(lycine) | Lys | K | 146.19 | ||
甲硫氨酸(methionine) | Met | M | 149.21 | ||
苯丙氨酸(phenylanaline) | Phe | P | 165.19 | ||
脯氨酸(proline) | Pro | P | 115.13 | ||
丝氨酸(serine) | Ser | S | 105.06 | ||
苏氨酸(threonine) | Thr | T | 119.12 | ||
色氨酸(tryptophan) | Trp | W | 204.22 | ||
酪氨酸(tyrosine) | Tyr | Y | 181.19 | 10.07 | |
缬氨酸(valine) | Val | P | 117.15 |
八、遗传密码
密码子的第二位
密码子的第一位(5’端) |
U |
C |
A |
G |
密码子的第三位(3端’) | ||||||
U |
UUU | Phe | UCU | Ser | UAU | Tyr | UGU | Gys | U | ||
UUC | Phe | UCC | Ser | UAC | Tyr | UGG | Gys | C | |||
UUA | Leu | UCA | Ser | UAA | 终止(赭石) | UGA | 终止(乳白) | A | |||
UUG | Leu | UGG | Ser | UAG | 终止(琥珀) | UGG | Trp | G | |||
C |
CUU | Leu | CCU | Pro | CAU | His | CGA | Arg | U | ||
CUC | Leu | CCC | Pro | CAC | His | CGC | Arg | C | |||
CUA | Leu | CCA | Pro | CAA | Gln | CGA | Arg | A | |||
CUG | Leu | CCG | Pro | CAG | Gln | CGC | Arg | G | |||
A |
AUU | Ile | ACU | Thr | AAU | Asn | AGU | Ser | U | ||
AUC | Ile | ACC | Thr | AAC | Asn | AGC | Ser | C | |||
AUA | Ile | ACA | Thr | AAA | Lys | AGA | Arg | A | |||
AUG | Met | ACG | Thr | AAG | Lys | AGC | Arg | G | |||
G |
GUU | Val | GCU | Ala | GAU | Asp | GGU | Gly | U | ||
GUC | Val | GCC | Ala | GAC | Asp | GGC | Gly | C | |||
GUA | Val | GCA | Ala | GAA | Glu | GGA | Gly | A | |||
GUG | Val | GCG | Ala | GAG | Glu | GGG | Gly | G |
九、常用酸碱技术参数
1.常见的市售酸碱的浓度
溶质 | 分子式 | 分子量 | mol/L | g/L | 重量(%) | 比重 | 配制mol/L溶液的加入量(ml/L) |
冰乙酸 | CH3COOH | 60.05 | 17.40 | 1045 | 99.5 | 1.050 | 57.5 |
乙酸 | 60.05 | 6.27 | 376 | 36 | 1.045 | 159.5 | |
甲酸 | HCOOH | 46.02 | 23.40 | 1080 | 90 | 1.200 | 42.7 |
盐酸 | HCl | 36.50 | 11.60 | 424 | 36 | 1.180 | 86.2 |
2.90 | 105 | 10 | 1.050 | 344.8 | |||
硝酸 | HNO3 | 63.02 | 15.99 | 1008 | 71 | 1.420 | 62.5 |
14.90 | 938 | 67 | 1.400 | 67.1 | |||
13.30 | 837 | 61 | 1.370 | 75.2 | |||
高氯酸 | HclO3 | 100.50 | 11.65 | 1172 | 70 | 1.670 | 85.8 |
9.20 | 923 | 60 | 1.540 | 108.7 | |||
磷酸 | H2PO4 | 80.00 | 18.10 | 1445 | 85 | 1.700 | 55.2 |
硫酸 | H2P`O4 | 98.10 | 18.00 | 1776 | 96 | 1.840 | 55.6 |
氢氧化铵 | NH4OH | 35.00 | 14.80 | 251 | 28 | 0.898 | 67.6 |
氢氧化钾 | KOH | 56.10 | 13.50 | 757 | 50 | 1.520 | 74.1 |
1.94 | 109 | 10 | 1.090 | 515.5 | |||
氢氧化钠 | NaOH | 40.00 | 19.10 | 763 | 50 | 1.530 | 52.4 |
2.75 | 111 | 10 | 1.110 | 363.4 |
2.各种浓度的酸碱贮存液的近似pH值
溶质 | 1Na | 0.1Na | 0.01Na | 0.001Na |
乙酸 | 0.40 | 2.90 | 3.40 | 3.90 |
盐酸 | 0.10 | 1.07 | 2.02 | 3.01 |
硫酸 | 0.30 | 1.20 | 2.10 | |
柠檬酸 | 2.10 | 2.60 | ||
氢氧化铵 | 11.80 | 11.30 | 10.80 | 10.30 |
氢氧化钠 | 14.05 | 13.07 | 12.12 | 11.13 |
碳酸氢钠 | 8.40 | |||
碳酸钠 | 11.50 | 11.00 |
J a.N为光量浓度[1N≈(1mol/L)×离子价数]。