Lab Tools - TanTec Biosystems

TanTec Biosystems, your source of viral reagents and tissue products

Compound	Formula	Molecular Weight	Specific Gravity	% by Weight	Conc. Reagent Molarity
Acetic acid, glacial	CH₃COOH	60.0	1.05	99.5	17.4
Formic acid	HCOOH	46.0	1.20	90	23.4
Hydrochloric acid	HCl	36.5	1.18	36	11.6
Nitric acid	HNO₃	63.0	1.42	71	16.0
Perchloric acid	HClO₄	100.5	1.67	70	11.6
Phosphoric acid	H₃PO₄	98.0	1.70	85	18.1
Sulfuric acid	H₂SO₄	98.1	1.84	96	18.0
Ammonium hydroxide	NH₄OH	35.0	0.90	28	14.8
Potassium hydroxide	KOH	56.1	1.52	50	13.5
Sodium hydroxide	NaOH	40.0	1.53	50	19.1
b-mercaptoethanol	HSCH₂CH₂OH	78.1	1.11	100	14.3

Buffers

Percentage by weight (w/v) = (% buffer desired/100) x final buffer vol (ml) = g of starting material needed.

Molar Solutions = desired molarity x formula weight x solution final volume (L) = grams needed

Henderson-Hasselbach Equation pH= pKa + log ([A-] / [HA])

Useful pH Ranges of Selected Biological Buffers (25 °C, 0.1M)

Buffers	Useful pH Range	pKa (20 ^oC)	pKa (25 ^oC)	pKa (37 ^oC)
MES	5.5–6.7	6.16	6.10	5.97
Bis-Tris	5.8–7.2	n/a	6.50	6.36
ADA	6.0–7.2	6.65	6.59	6.46
Aces	6.1–7.5	6.88	6.78	6.54
PIPES	6.1–7.5	6.80	6.76	6.66
MOPSO	6.2–7.6	n/a	6.90	6.75
Bis-Tris Propane	6.3–9.5	n/a	6.8, 9.0	n/a
BES	6.4–7.8	7.17	7.09	6.90
MOPS	6.5–7.9	7.28	7.20	7.02
TES	6.8–8.2	7.50	7.40	7.16
HEPES	6.8–8.2	7.55	7.48	7.31
DIPSO	7.0–8.2	n/a	7.60	7.35
MOBS	6.9–8.3	n/a	7.60	n/a
TAPSO	7.0–8.2	n/a	7.60	7.39
Trizma	7.0–9.0	8.20	8.06	7.72
HEPPSO	7.1–8.5	n/a	7.80	6.66
POPSO	7.2–8.5	n/a	7.80	7.63
TEA	7.3–8.3	n/a	7.80	n/a
EPPS	7.3–8.7	n/a	8.00	n/a
Tricine	7.4–8.8	8.16	8.05	7.80
Gly-Gly	7.5–8.9	n/a	8.20	n/a
Bicine	7.6–9.0	8.35	8.26	8.04
HEPBS	7.6–9.0	n/a	8.30	n/a
TAPS	7.7-9.1	8.49	8.40	8.18
AMPD	7.8–9.7	n/a	8.80	n/a
TABS	8.2–9.6	n/a	8.90	n/a
AMPSO	8.3–9.7	n/a	9.00	9.10
CHES	8.6–10.0	9.55	9.49	9.36
CAPSO	8.9–10.3	n/a	9.60	9.43
AMP	9.0–10.5	n/a	9.70	n/a
CAPS	9.7–11.1	10.56	10.40	10.02
CABS	10.0–11.4	n/a	10.70	n/a

Tris Buffer Table – pH vs Temperature

pH at Temperature			g/L for 0.05 M Solution
5 °C	25 °C	37 °C	Tris-HCL	Tris-Base
7.76	7.20	6.91	7.02	0.67
7.89	7.30	7.02	6.85	0.80
7.97	7.40	7.12	6.61	0.97
8.07	7.50	7.22	6.35	1.18
8.18	7.60	7.30	6.06	1.39
8.26	7.70	7.40	5.72	1.66
8.37	7.80	7.52	5.32	1.97
8.48	7.90	7.62	4.88	2.30
8.58	8.00	7.71	4.44	2.65
8.68	8.10	7.80	4.02	2.97
8.78	8.20	7.91	3.54	3.34
8.88	8.30	8.01	3.07	3.70
8.98	8.40	8.10	2.64	4.03
9.09	8.50	8.22	2.21	4.36
9.18	8.60	8.31	1.83	4.65
9.28	8.70	8.42	1.50	4.90
9.36	8.80	8.51	1.23	5.13
9.47	8.90	8.62	0.96	5.32
9.56	9.00	8.70	0.76	5.47

Phosphate Buffer Table – 0.2M solution

Na-Phosphate Monobasic Anhydrous (g/L)	Na-Phosphate Dibasic Heptahydrate (g/L)	pH 23 °C
22.4	3.49	5.7
22.08	4.29	5.8
21.60	5.37	5.9
21.05	6.60	6.0
20.40	8.05	6.1
19.56	9.93	6.2
18.60	12.07	6.3
17.64	14.22	6.4
16.44	16.90	6.5
15.00	20.12	6.6
13.56	23.34	6.7
12.24	26.29	6.8
10.80	29.51	6.9
9.36	32.73	7.0
7.92	35.95	7.1
6.72	38.63	7.2
5.52	41.31	7.3
4.56	43.46	7.4
3.84	45.07	7.5
3.12	46.68	7.6
2.52	48.55	7.7
2.04	49.09	7.8
1.68	49.89	7.9
1.27	50.81	8.0

Citric Acid – Na₂HPO₄ Buffer Solutions, pH approx. 2.6–7.61

Citric acid monohydrate, C₆H₈O₇ H₂O, M. wt. 210.14

0.1M-solution contains 21.01 g/l

Na₂HPO₄, M. wt. 141.98;0.2M-solution contains 28.40 g/l

Na₂HPO4 2H₂O, M. wt. 178.05; 0.2M-solution contains 35.61 g/l

x ml 0.1M-citric acid and y ml 0.2M-Na2HPO4 mixed

pH	x ml 0.1M-citric acid	y ml 0.2-Na₂HPO₄
2.6	89.10	10.90
2.8	84.15	15.85
3.0	79.45	20.55
3.2	75.30	24.70
3.4	71.50	28.50
3.6	67.80	32.20
3.8	64.50	35.50
4.0	61.45	38.55
4.2	58.60	41.40
4.4	55.90	44.10
4.6	53.25	46.75
4.8	50.70	49.30
5.0	48.50	51.50
5.2	46.40	53.60
5.4	44.25	55.75
5.6	42.00	58.00
5.8	39.55	60.45
6.0	36.85	63.15
6.2	33.90	66.10
6.4	30.75	69.25
6.6	27.25	72.75
6.8	22.75	77.25
7.0	17.65	82.35
7.2	13.05	86.95
7.4	9.15	90.85
7.6	6.35	93.65

Citric Acid – Sodium Citrate Buffer Solutions, pH 3.0–6.21

Citric acid monohydrate, C₆H₈O₇ H2O, M. wt. 210.14

0.1M-solution contains 21.01 g/l

Trisodium citrate dihydrate, C₆H₅O₇Na₃ 2H₂O, M. wt. 294.12

0.1M-solution contains 29.41 g/l

x ml 0.1M-citric acid

y ml 0.1M-trisodium citrate mixed

pH	x ml 0.1 M-citric acid	y ml 0.1M-trisodium citrate
3.0	82.0	18.0
3.2	77.5	22.5
3.4	73.0	27.0
3.6	68.5	31.5
3.8	63.5	36.5
4.0	59.0	41.0
4.2	54.0	46.0
4.4	49.5	50.5
4.6	44.5	55.5
4.8	40.0	60.0
5.0	35.0	65.0
5.2	30.5	69.5
5.4	25.5	74.5
5.6	21.0	79.0
5.8	16.0	84.0
6.0	11.5	88.5
6.2	8.0	92.0

Sodium Acetate – Acetic Acid Buffer Solutions, pH 3.7–5.61

Sodium acetate trihydrate, CH₃COONa 3H₂O, M. wt. 136.09

0.2M-solution contains 27.22 g/l

x ml 0.2M-NaOAc

y ml 0.2M-HOAc mixed

pH, 18 °C	x ml 0.2M-NaOAc	y ml 0.2M-HOAc
3.7	10.0	90.0
3.8	12.0	88.0
4.0	18.0	82.0
4.2	26.5	73.5
4.4	37.0	63.0
4.6	49.0	51.0
4.8	59.0	41.0
5.0	70.0	30.0
5.2	79.0	21.0
5.4	86.0	14.0
5.6	91.0	9.0

Na₂HPO₄ – NaH₂PO₄ Buffer Solutions, pH 5.8–8.0 at 25 °C

Na₂HPO₄ 2H₂O, M. wt. 178.05

0.2M-solution contains 35.61 g/l, Na₂HPO₄ 12H₂O, M. wt. 358.22

0.2M -solution contains 71.64 g/l, NaH₂PO₄ H₂O, M. wt. 138.01

0.2M-solution contains 27.6 g/l, NaH₂PO₄ 2H₂O, M. wt. 156.03

0.2M-solution contains 31.21 g/l

x ml 0.2M-Na2HPO4, y ml 0.2M-NaH₂PO₄; diluted to 100 ml with H₂O

pH, 25 °C	x ml 0.2M-Na₂HPO₄	y ml 0.2M-NaH₂PO₂
5.8	4.0	46.0
6.0	6.15	43.85
6.2	9.25	40.75
6.4	13.25	36.75
6.6	18.75	31.25
6.8	24.5	25.5
7.0	30.5	19.5
7.2	36.0	14.0
7.4	40.5	9.5
7.6	43.5	6.5
7.8	45.75	4.25
8.0	47.35	2.65

Imidazole (glyoxaline) – HCl buffer solutions, pH 6.2–7.8 at 25 °C

Imidazole, C3H4N2, M. wt. 68.08

25 ml 0.2M-imidazole (13.62 g/l), x ml 0.2M-HCl, diluted to 100 ml with H₂O

pH, 25 °C	x ml 0.2M-HCl
6.2	21.45
6.4	19.9
6.6	17.75
6.8	15.2
7.0	12.15
7.2	9.3
7.4	6.8
7.6	4.65
7.8	3.0

Sodium Carbonate – Sodium Bicarbonate Buffer Solutions, pH 9.2–10.81

Na₂CO₃ 10H₂O, M. wt. 286.2; 0.1M-solution contains 28.62 g/l

NaHCO₃, M. wt. 84.0; 0.1M-solution contains 8.40 g/l

x ml 0.1M-Na₂CO₃ and y ml 0.1M-Na₂HCO₃ mixed

pH	x ml 0.1M-Na₂CO₃	y ml 0.1M-NaHCO₃
20 °C	37 °C
9.2	8.8	10	90
9.4	9.1	20	80
9.5	9.4	30	70
9.8	9.5	40	60
9.9	9.7	50	50
10.1	9.9	60	40
10.3	10.1	70	30
10.5	10.3	80	20
10.8	10.6	90	10

Electrophoresis

% Gel	Optimum resolution for linear DNA (kb)
0.5	30 to 1.0
0.7	12 to 0.8
1.0	10 to 0.5
1.2	7 to 0.4
1.5	3 to 0.2

DNA facts

Average weight of a DNA base pair (sodium salt) = 650 daltons

A₂₆₀ unit ds DNA = 50 mg/ml = 0.15 mM (in nucleotides)

A₂₆₀ unit ss DNA = 33 mg/ml = 0.10 mM (in nucleotides)

A₂₆₀ unit ss RNA = 40 mg/ml = 0.11 mM (in nucleotides)

MW of a ds DNA molecule = (# of base pairs) X (650 daltons/base pair)

Moles of ends of a ds DNA molecule = 2 X (grams of DNA) ÷ MW in daltons)

Moles of ends generated by restriction endonuclease cleavage:

circular DNA molecule: 2X (moles of DNA) X (# of sites)

linear DNA molecule: 2X (moles of DNA) X (# of sites) ÷ 2 X (moles of DNA)1 g of

1 mg of 1000 bp DNA = 1.52 pmol = 9.1x10¹¹ molecules

1 mg of puc18/19 (2686 bp) = 0.57 pmol = 3.4 x 10¹¹ molecules

1 mg of lambda DNA (48502 bp) = 0.33 pmol = 1.8 x 10¹⁰ molecules

1 pmol of 1000 bp of DNA = 0.66 g

1 pmol of puc18/19 DNA (2686 bp) = 1.77 g

1 pmol of DNA (48502 bp) = 32.01 g

pmol/100 µg DNA = 100 µg DNA x pmol/660 (~ mw of a base pair) x 10⁶pg/1 ug x 1/genome size (bp)

Genome/100 g = pmol/100 g * 6.022 × 10²³(Avogadro’s number)

Protein facts

Codon table

		2nd base
		U	C	A	G
1st base	U	UUU Phenylalanine UUC Phenylalanine UUA Leucine UUG Leucine	UCU Serine UCC Serine UCA Serine UCG Serine	UAU Tyrosine UAC Tyrosine UAA Ochre (Stop) UAG Amber (Stop)	UGU Cysteine UGC Cysteine UGA Opal (Stop) UGG Tryptophan
	C	CUU Leucine CUC Leucine CUA Leucine CUG Leucine	CCU Proline CCC Proline CCA Proline CCG Proline	CAU Histidine CAC Histidine CAA Glutamine CAG Glutamine	CGU Arginine CGC Arginine CGA Arginine CGG Arginine
	A	AUU Isoleucine AUC Isoleucine AUA Isoleucine AUG Methionine	ACU Threonine ACC Threonine ACA Threonine ACG Threonine	AAU Asparagine AAC Asparagine AAA Lysine AAG Lysine	AGU Serine AGC Serine AGA Arginine AGG Arginine
	G	GUU Valine GUC Valine GUA Valine GUG Valine	GCU Alanine GCC Alanine GCA Alanine GCG Alanine	GAU Aspartic acid GAC Aspartic acid GAA Glutamic acid GAG Glutamic acid	GGU Glycine GGC Glycine GGA Glycine GGG Glycine

The codon AUG both codes for methionine and serves as an initiation site: the first AUG in an mRNA's coding region is where translation into protein begins.