Bio-chemical laboratory commonly used technical parameters data

, nucleic acids and proteins
common

1. The physical constant of nucleoside triphosphate
compound molecular weight 1 molar solution (pH7.0) OD280/OD260
ATP 507 259 15400 0.15
CTP 483 271 9000 0.97
GTP 523 253 13700 0.66
UTP 484 262 10000 0.38
dATP 494 259 15200 0.15
dCTP 467 271 93 00 0.98
dGTP 507 253 13700 0.66
dTTP 482 267 9600 0.71
2. Length and molecular weight of common nucleic acids
Nucleotides Molecular weight
-DNA 48502 (double-stranded ring) 3.0×107
pBR322 4 363 (double chain) 2.8×106
28SrRNA 4800 1.6×106
23SrRNA 3700 1.2×106
18SrRNA 1900 6.1×105
19SrRNA 1700 5.5× (105
5SrRNA 120 3.6×104
tRNA (E. coli) 75 2.5×104
3. Common nucleic acid protein conversion data
(1) Weight conversion
1 sg s 10-6g1pg s 10-12g
1ng s 10-9g1fg s 10-15g
(2) points Photomethic conversion:
1A260 double-stranded DNA: 50 sg/ml
1A260 single-stranded DNA s 30 sg/ml
1A260 single-stranded RNA s 40 sg/ml
(3) DNA Molar conversion:
1 μg 100bp DNA=1.52pmol=3.03pmol End
1μg pBR322 DNA=0.36pmol
1pmol 1000bp DNA=0.2 66
1pmol pBR322 s 2.8 sg
1kb double-stranded DNA (sodium salt) s6.6×105 Dalton
1kb single-stranded DNA (sodium salt) s 3.3× 105 Dalton
1kb single-stranded RNA (sodium salt) s3.4×105 Dalton
(4) protein molar conversion:
100pmol molecular weight 100,000 protein s1 50,000 proteins
100pmol molecular weight of 0 100pmol , 5 g
100pmol molecular weight 10,000 proteins , 1 mg
amino acids average molecular weight of 126.7 Dalton
(5) Protein/DNA Conversion:
1kb DNA s 333 amino acid coding capacity s 3.7×104MW protein
10,000MW protein s 270bp DNA
30,000MW Protein - 810bp DNA
50,000MW Protein - 1.35kb
100,000MW Protein - 2.7kb DNA
4. Common protein molecular weight standard reference
(1) high molecular weight standard reference (2) molecular weight standard reference (3) low molecular weight standard reference
myoprotein molecular weight phosphatase B 97,400 carbonate enzyme 31,00
myococrine 212,00 0
serum
albumin 66,200 soyprotease 21,500
β-semi-lactose glycase B 116,000 glutamate dehydrogenase 55,000 inhibitors
phosphoric acid Enzyme B 97,400 albumin 42,700 horse heart myoglobulin 16,900
bovine serum albumin 66,200 aldehyde shrinkase 40,000 lysolyzyme 14,400
hydrohydrolysisase' 57,000 carbon anhydrase 31,000 creatinine (F1) 8,100
aldehyde shrinkase 40,000 soyprotease 21,500 tglobulin (F2) 6,200
Inhibitor myoglobulin (F3) 2,500
14,400
5. Common standard references for DNA molecular weight
-DNA/HindIII. S.DNA/EcoRI. s/Hind III. EcoRI. pBR322/HaeIII.
23130 21226 21227 587 123
9416 7421 5148 405 104
6557 5804 49 73 504 89
4361 5643 4268 458 80
2322 4843 3530 434 64
2027 3 530 2027 267 57
5641904 234 51
1251584 213 21
1375 192 18
97 4 184 11
831 124 7
564
125
Continue to Table
pBR322/HinfI. φχ174/HinfI. φχ174 /Hae III. φχ174/TapI.
1631 726 140 1353 2914
517 713 118 1078 1175
506 55 3 100 872 404
396 500 82 603 327
344 417 66 310 231


298 413 48 281 141
221 311 42 271 87
220 249 40 234 54
154 200 24 194 33
75 151118 20
72
II, Common buffer
1. Molecular cloning commonly used buffer
2. Preparation of potassium phosphate buffer at
(1)25 degrees C
pH 1mol/L K2HPO4 (ml) 1mol/L KH2PO4 (ml)
5.8 8.5 91.5
(6.0) 13.2 86.8
6.2 19.2 80.8
6.4 27.8 72.2
6.6 38.1 61.9
6.8 49.6 7 50.3
7.0 61.5 38.5
7.2 71.7 28.3
7.4 80.2 19.8
7.6 86.6 13.4
7.8 90.8 9.2
8.0 94.0 6.2(2)25 degrees C at 0.1mol/L sodium phosphate buffer preparation s
pH 1mol/L Na2HPO4 (ml) 1mo l/L NaH2PO4 (ml)
5.8 7.9 92.1
6.0 12.0 88.0
6.2 17.8 82.2
6.4 25.5 74. 5
6.6 35.2 64.8
6.8 46.3 53.7
7.0 57.7 42.3
7.2 68.4 31.6
7.4 77.4 22.6
7.6 84.5 15.5
7.8 89.6 10.4
8.0 93.2 6.8
: Mixed with distilled water The two 1mol/L storage fluids are diluted to 1000 ml and their pH is calculated according to the Henderson-Hasselbalch equation:
pH-pK' 1g (proton receptors)/(proton feeders))
here, pK'-6.86 (25 degrees C).
3. Electrophoresis buffer
sequencing gel dosing buffer
98% deionized methamide
10mol/L EDTA (pH8.0)
0.025% Xylene FF
0.025% bromophenol blue
methylamide: many lot numbers of
-reagents
-grade methamphetamine, its purity meets the requirements of use, no further treatment. However, once slightly yellow, it is applied to the magnetic mixer to mix methamide and Dowex XG8 mixing bed resin for 1 hour for deionization treatment, and with Whatman 1 filter paper
filtration
2 times, deionized methamide into small parts, nitrogen charged at -70 degrees C.
commonly used electrophoretic buffer
buffer Using liquid thick storage fluid (per liter)
Tris-acetic acid (TAE) 1×:0.04mol/L Tris-acetic acid 50×:242g Tris alkali
0 .001mol/L EDTA 57.1ml ice acetic acid
100ml 0.5mol/L EDTA (pH8.0)
Tris-phosphoric acid (TPE) 1×:0.09mol/L Tris-Phosphoric acid 10×:10g Tris alkali
0.002mol/L EDTA 15.5ml 85% phosphoric acid (1.679g/ml)
40ml 0.5mol/L EDTA (pH8. 0)
Tris-boric acid (TBE) a 0.5×0.045mol/L Tris-boric acid 5×:54g Tris alkali
0.001mol/L EDTA 27.5 boric acid
20ml 0.5mol/L EDTA (pH8.0)
alkaline bufferb 1×:50mmol/L NaOH 1×:5ml 10mol/L NaOH
1mmol/L EDTA 2ml 0.5mmol/L EDTA (pH8.0)
Tris-Glyc 1×:25mmol/L Tris 5 ×: 15.1g Tris
250mmol/L glycine 94g glycine (electrophoretic grade) (pH8.3)
0.1% SDS 50ml 10 % SDS (electrophoresis grade)
Description:
(1) TBE solution after a long period of storage will form sediment, in order to avoid this problem, can be stored at room temperature with a glass bottle 5× solution, after precipitation is discarded.
are electrophoresis with 1×TBE as a liquid (i.e., 1:5 dilution of
storage
agarose gel). However, 0.5 × the liquid used in the water has sufficient buffer capacity. At present, almost all agar gum electrophoresis uses 1:10 diluted storage fluid as a liquid for use.
buffer tank for polyacrylamide gel vertical grooves is small, so the flow through the buffer is usually large and requires 1×TBE to provide sufficient buffer capacity.
(2) alkaline electrophoresis buffer should be rinsed.
(3) Tris-Glycine buffer electrophoresis with SDS polyacrylamide gel.
2×SDS gel dosing buffer:
100mmol/L Tris s #8226; HCl (6.8)
200mmol/L dithionol (DTT)
4% SDS (DTT) Electrophoresis grade)
0.2% bromophenol blue
20% glycelin
DTT-free 2×SDS gel sample buffer can be stored at room temperature, should be taken before use 1mol / L storage liquid is added to the above buffer.
4. Gel sample buffer
buffer type 6× buffer storage temperature I. 0.25% bromophenol blue
4 degrees C
0.25% xylene green FF
40% (W/V) sucrose water solution <br/